Selective androgen receptor modulators (SARMs) and uses thereof

ABSTRACT

Provided herein are compounds, such as compounds of Formula I, that bind to androgen receptors and/or modulate activity of androgen receptors. Also provided are methods for making and using such compounds. Also provided are compositions including such compounds and methods for making and using such compositions.

RELATED APPLICATIONS

This application is a divisional of co-pending allowed U.S. patentapplication Ser. No. 12/734,993, filed Jun. 7, 2010, which is the U.S.National Stage application of PCT/US2008/013657, filed Dec. 12, 2008,and entitled “SELECTIVE ANDROGEN RECEPTOR MODULATORS (SARMs) AND USESTHEREOF,” which claims benefit of priority to U.S. Provisional PatentApplication Ser. No. 61/008,731, to Lin Zhi, filed on Dec. 21, 2007,entitled “SELECTIVE ANDROGEN RECEPTOR MODULATORS (SARMs) AND USESTHEREOF.” Where permitted, the subject matter of each of the-abovementioned applications is incorporated by reference in its entirety.

FIELD

Provided herein are selective androgen receptor modulator (SARM)compounds that bind to androgen receptors and/or modulate activity ofandrogen receptors, and to methods for making and using such compounds.Also provided are compositions including such compounds and methods formaking and using such compositions. Also provided are methods for thetreatment of androgen receptor mediated diseases.

BACKGROUND

Certain intracellular receptors (IRs) have been shown to regulatetranscription of certain genes (e.g., see R. M. Evans, Science 240: 889(1988)). Certain of such IRs are steroid receptors, such as androgenreceptors, estrogen receptors, mineralo-corticoid receptors, andprogesterone receptors. Gene regulation by such receptors typicallyinvolves binding of an IR by a ligand.

In certain instances, a ligand binds to an IR, forming a receptor/ligandcomplex. Such a receptor/ligand complex can then translocate to thenucleus of a cell, where it binds to the DNA of one or more generegulatory regions. Once bound to the DNA of a particular generegulatory region, a receptor/ligand complex can modulate the productionof the protein encoded by that particular gene. In certain instances, anandrogen receptor/ligand complex regulates expression of certainproteins. In certain instances, an androgen receptor/ligand complex caninteract directly with the DNA of a particular gene regulatory region orwith other transcription factors. In certain instances, suchinteractions result in modulation of transcriptional activation.

Androgen therapy has been used to treat a variety of male disorders suchas reproductive disorders and primary or secondary male hypogonadism. Anumber of natural or synthetic AR agonists have been investigated forthe treatment of musculoskeletal disorders, such as bone disease,hematopoietic disorders, neuromuscular disease, rheumatological disease,wasting disease, and for hormone replacement therapy (HRT), such asfemale androgen deficiency. In addition, AR antagonists, such asflutamide and bicalutamide, are used to treat prostate cancer. Theeffectiveness of known modulators of steroid receptors is often temperedby their undesired side-effect profile, particularly during long-termadministration. For example, potential side effects of androgen therapyfor women include acne, weight gain, excess facial and body hair,permanent lowering of the voice, and adverse lipid changes. In men,adverse effects can include disordered sleep and breathing,polycythemia, and repression of high density lipoprotein. Thus there isa need for compounds that do not exhibit the adverse side-effects. It isamong the objects herein to provide such compounds that modulate theactivity of androgen receptor.

SUMMARY

Compounds for use in compositions and methods for modulating theactivity of androgen receptor are provided. The compounds providedherein are non-steroidal Selective Androgen Receptor Modulators orSARMs. In particular, non-steroidal SARMs display therapeutic benefitbut generally do not display adverse androgenic effects, such asprostate enlargement, acne, hirsutism, virilization and masculinization.The compounds selectively modulate (agonize or antagonize) the functionof the AR, such as in a tissue-selective manner, to produce the effectsof androgens without or with reduced negative or undesired androgenicproperties. Among the compounds provided herein are agonists of androgenreceptor. Among the compounds provided herein are antagonists ofandrogen receptor. Among the compounds provided herein are androgenreceptor partial agonists.

Among the compounds provided herein are tissue specific selectiveandrogen receptor modulators. They can be used for oral testosteronereplacement therapy. Compounds provided herein display agonist activitywith EC₅₀ values generally less than 1 micromolar. Compounds providedherein display antagonist activity with IC₅₀ values generally less than2 micromolar. SARMs provided herein generally target anabolic tissue,such as connective tissue, including bone and muscle, and can be used toincrease the mass of a connective tissue in a subject and to reverseconnective tissue loss in a subject. Among the disorders that can betreated are muscle wasting, cachexia, frailty and osteoporosis and othermuscle and bone disorders, including those enumerated below.

Compounds provided herein have a structure of Formula I or Formula II orFormula III:

where R¹ is halogen, pseudohalogen, optionally substituted lower alkyl,optionally substituted haloalkyl or NO₂, particularly lower haloalkyl orhalogen, and in particular is CF₃, F, or Cl; R² is hydrogen, halogen,pseudohalogen, optionally substituted lower alkyl or optionallysubstituted lower haloalkyl, particularly hydrogen or methyl; R³ ishydrogen, halogen, pseudohalogen, optionally substituted lower alkyl oroptionally substituted lower haloalkyl, particularly hydrogen or loweralkyl, and in particular hydrogen or methyl; R⁴ is halogen or lowerhaloalkyl, particularly CF₃ or halogen, and in particular Cl or CF₃; andR⁵ is lower alkyl or lower haloalkyl, particularly C₁ to C₄ alkyl or C₁to C₄ haloalkyl, and in particular methyl, ethyl or CF₃. Also providedare pharmaceutically acceptable salts, esters and prodrugs of compoundsof Formula I or Formula II or Formula III.

In some embodiments, the compounds provided herein exhibit tissueselective androgen receptor agonist activity. In some embodiments, thecompounds provided herein exhibit tissue selective androgen receptorantagonist activity. In some embodiments, the compounds provided hereinare androgen receptor selective binding compounds.

Compounds provided herein are effective for treating one or moreandrogen receptor mediated diseases or conditions. Such conditions anddiseases include those caused by androgen deficiency and/or those thatcan be ameliorated by androgen administration. In certain embodiments,compounds provided herein are effective for treating one or morediseases or conditions responsive to an androgen receptor agonist. Incertain embodiments, compounds provided herein are effective in treatingone or more conditions whose etiology involves hypoactivity orsubsensitivity of androgen receptor. In other embodiments, compoundsprovided herein are effective for treating one or more diseases orconditions responsive to an androgen receptor antagonist. In otherembodiments, compounds provided herein are effective in treating one ormore conditions whose etiology involves hyperactivity of androgenreceptor.

In some tissues, the compounds provided herein can exhibit AR agonistactivity and can be used to treat conditions that are caused by androgendeficiency or hypoactivity or subsensitivity of androgen receptor, orthat can be ameliorated by androgen replacement or are responsive totreatment with an AR agonist. Such conditions, include, but not limitedto, aging skin; Alzheimer's disease; anemias, such as for example,aplastic anemia; anorexia; arthritis, including inflammatory arthritis,rheumatoid arthritis, osteoarthritis and gout; arteriosclerosis;atherosclerosis; bone disease, including metastatic bone disease; bonedamage or fracture, such as by accelerating bone fracture repair and/orstimulation of osteoblasts and/or stimulation of bone remodeling and/orstimulation of cartilage growth; distraction osteogenesis; reduced bonemass, density or growth; bone weakening, such as induced byglucocorticoid administration; musculoskeletal impairment (e.g., in theelderly); cachexia; cancer, including breast cancer and osteosarcoma;cardiac dysfunction (e.g., associated with valvular disease, myocardialinfarction, cardiac hypertrophy or congestive heart failure);cardiomyopathy; catabolic side effects of glucocorticoids; Crohn'sdisease; growth retardation in connection with Crohn's disease; shortbowel syndrome; irritable bowel syndrome; inflammatory bowel disease;ulcerative colitis; cognitive decline and impairment; dementia; shortterm memory loss; contraception (male and female); chronic obstructivepulmonary disease (COPD); chronic bronchitis; decreased pulmonaryfunction; emphysema; decreased libido in both men and women; depression;nervousness, irritability and/or stress; reduced mental energy and lowself-esteem (e.g., motivation/assertiveness); dyslipidemia; erectiledysfunction; frailty; age-related functional decline (“ARFD”) in theelderly; growth hormone deficiency; hematopoietic disorders; hormonereplacement (male and female); hypercholesterolemia; hyperinsulinemia;hyperlipidemia; hypertension; hyperandrogenemia; hypogonadism (includingprimary and secondary); hypothermia (including hypothermia followinganesthesia); impotence; insulin resistance; type 2 diabetes;lipodystrophy (including in subjects taking HIV or AIDS therapies suchas protease inhibitors); male menopause; metabolic syndrome (syndromeX); loss of muscle strength and/or function (e.g., in the elderly);muscular dystrophies; muscle loss following surgery (e.g., post-surgicalrehabilitation); muscular atrophy (e.g., due to physical inactivity, bedrest or reduced weight-bearing conditions such as microgravity);neurodegenerative diseases; neuromuscular disease; decreased plateletcount; platelet aggregation disorders; obesity; osteoporosis;osteopenia; glucocorticoid-induced osteoporosis;osteochondro-dysplasias; periodontal disease; premenstrual syndrome;postmenopausal symptoms in women; Reaven's syndrome; rheumatologicaldisease; sarcopenia; male and female sexual dysfunction (e.g., erectiledysfunction, decreased sex drive, sexual well-being, decreased libido);physiological short stature, including growth hormone deficient childrenand short stature associated with chronic illness and growth retardationassociated with obesity; tooth damage (such as by acceleration of toothrepair or growth); thrombocytopenia; vaginal dryness; atrophicvaginitis; ventricular dysfunction; wasting, including wasting secondaryto fractures and wasting in connection with chronic obstructivepulmonary disease (COPD), chronic liver disease, AIDS, weightlessness,cancer cachexia, burn and trauma recovery, chronic catabolic state(e.g., coma), eating disorders (e.g., anorexia), chemotherapy, multiplesclerosis or other neurodegenerative disorders.

In some tissues, the compounds provided herein exhibit AR agonistactivity and can be used to stimulate pulsatile growth hormone release;in hormone replacement therapy, such as female androgen deficiency andmale androgen decline; to improve bone strength, muscle strength andtone; to reduce subcutaneous fat in a subject; to enhance bone andmuscle performance/strength; to increase athletic performance; toattenuate or reverse protein catabolic responses following trauma (e.g.,reversal of the catabolic state associated with surgery, congestiveheart failure, cardiac myopathy, burns, cancer, COPD); to improve sleepquality and/or correct the relative hyposomatotropism of senescence dueto high increase in REM sleep and a decrease in REM latency; and totreat age related decreased testosterone levels in men.

In some tissues, the compounds provided herein can exhibit AR antagonistactivity and can be used to treat conditions whose etiology involveshyperactivity of androgen receptor or that are responsive to treatmentwith an AR antagonist. Such conditions include, but are not limited to,acanthosis nigricans, acne, adrenal hyper-androgenism, androgeneticalopecia (male-pattern baldness), adenomas and neoplasias of theprostate (e.g., advanced metastatic prostate cancer), benign prostatehyperplasia, cancer (e.g., cancer of the breast, bladder, endometrium,lung (non-small cell lung cancer), pancreas, prostate, includingandrogen dependent prostate cancer, and skin); bulimia nervosa; chronicfatigue syndrome (CFS); chronic myalgia; acute fatigue syndrome;contraception; counteracting preeclampsia, eclampsia of pregnancy andpreterm labor; delayed wound healing; erythrocytosis; gestationaldiabetes; hirsutism; hyperinsulinemia including nesidioblastosis;hyperandrogenism; hypercortisolism; Cushing's syndrome; hyperpilosity;infertility; malignant tumor cells containing the androgen receptor,such as is the case for breast, brain, skin, ovarian, bladder,lymphatic, liver and kidney cancers; menstrual irregularity; ovarianhyperandrogenism; polycystic ovarian syndrome; seborrhea; sleepdisorders; sleep apnea; and visceral adiposity.

In certain embodiments, compounds provided herein are effective fortreating prostate cancer. In certain embodiments, compounds providedherein are effective for treating androgen dependant prostate cancer. Incertain embodiments, compounds provided herein are effective fortreating androgen independent prostate cancer. In certain embodiments,compounds provided herein are effective for treating androgenindependent androgen receptor dependent prostate cancer.

The methods of treatment are practiced by administering to the subject acompound provided herein. In certain embodiments, provided herein aremethods for treating a condition responsive to androgen receptormodulation in a subject by identifying a subject in need of suchtreatment and administering to the subject a compound provided herein.In certain embodiments, the methods provided herein are for treating adisease or condition responsive to an androgen receptor agonist. Incertain embodiments, the methods provided herein are for treating acondition responsive to an androgen receptor antagonist.

In certain embodiments, provided herein are methods for modulating anactivity of an androgen receptor by contacting an androgen receptor withat least one compound provided herein. In certain such embodiments, theandrogen receptor is in a cell. In some embodiments, the modulation isagonizing the receptor. In some embodiments, the modulation isantagonizing the receptor.

In certain embodiments, provided herein are methods for identifying acompound that is capable of modulating an activity of an androgenreceptor by contacting a cell expressing an androgen receptor with acompound provided herein and monitoring an effect of the compound uponthe cell.

In certain embodiments, provided herein are methods of contraception ina subject. The methods include administering to the subject a compoundof Formula I, II or III or pharmaceutically acceptable salts or prodrugsthereof, in an amount effective to provide contraception. In someembodiments, the compound is co-administered with an androgen selectedfrom among testosterone, 19-nortestosterone,7α-methyl-19-nortestosterone and 5α-dihydro-testosterone. In oneembodiment, the subject is male and a compound of formula I, II or IIIis administered in an amount effective to suppress sperm production inthe subject, thereby effecting contraception in the subject. In oneembodiment, the compounds provided herein inhibit spermatogenesis in asubject. In one embodiment, the subject is female and a compound offormula I, II or III is administered in an amount effective to providecontraception in the subject.

In certain embodiments, provided herein are methods for providinghormone therapy. The methods include administering to the subject acompound of formula I, II or III, in an amount effective to modulateandrogen receptor activity, and thereby effect a change in anandrogen-dependent condition. In some tissues, the compound is anandrogen receptor agonist. In some tissues, the compound is an androgenreceptor antagonist.

In certain embodiments, provided herein are methods for treating cancerin a subject, comprising administering to the subject a compound ofFormula I, II or III or pharmaceutically acceptable salts or prodrugsthereof, in an amount effective to treat cancer in the subject. Incertain embodiments, the cancer is selected from among breast cancer,colorectal cancer, gastric carcinoma, glioma, head and neck squamouscell carcinoma, skin cancer, papillary renal carcinoma, leukemia,lymphoma, Li-Fraumeni syndrome, malignant pleural mesothelioma,melanoma, multiple myeloma, non-small cell lung cancer, synovialsarcoma, thyroid carcinoma, transitional cell carcinoma of urinarybladder, and prostate cancer. In some embodiments, the compound isadministered in an amount effective to kill the cancerous cells. In someembodiments, the compound is administered in an amount effective toinhibit growth and/or metastasis of the cancer. In some embodiments, thecompound is co-administered with one or more other therapeutic agentsselected from among anti-proliferative agents, anti-tumor agents,adrenocorticosteroids, progestins, estrogens, antiestrogens,radionuclides, toxins and cytotoxic drugs, chemotherapy agents,photodynamic therapy dyes and antibiotics or combinations thereof.

In certain embodiments, provided herein are methods of treating prostatecancer in a subject. The methods include administering to the subject acompound of Formula or III or pharmaceutically acceptable salts orprodrugs thereof, in an amount effective to treat prostate cancer in thesubject. In some embodiments, the prostate cancer is androgen dependantprostate cancer. In some embodiments, the prostate cancer is androgenindependent prostate cancer. In some embodiments, the prostate cancer isandrogen independent, but androgen receptor dependant prostate cancer.In some embodiments, the compound is administered to the subject in anamount effective to kill the cancerous cells. In some embodiments, thecompound is administered to the subject in an amount effective toinhibit the growth and/or metastasis of the prostate cancer cells. Insome embodiments, the compound is co-administered with anothertherapeutic agent selected from among flutamide, a toxin, bicalutamide,nilutamide, an anti-tumor agent, a cytotoxic drug, a radio-nuclide andcombinations thereof. In some embodiments, the compound and/or anothertherapeutic agent, if present, is selectively targeted to react withprostate cancer cells by conjugating the compound and/or therapeuticagent to a prostate tumor antigen.

In certain embodiments, provided herein are methods of delaying theprogression of prostate cancer in a subject. The methods includeadministering to the subject a compound of Formula I, II or III orpharmaceutically acceptable salts or prodrugs thereof, in an amounteffective to delay the progression of prostate cancer in the subject.

In certain embodiments, provided herein are methods of improvingathletic performance in a subject. The methods include administering tothe subject a compound of Formula I, II or III or pharmaceuticallyacceptable salts or prodrugs thereof in an amount effective to improveathletic performance in the subject.

In certain embodiments, provided herein are methods of increasing muscleperformance, muscle size and/or muscle strength in a subject. Themethods include administering to the subject a compound of Formula I, IIor III or pharmaceutically acceptable salts or prodrugs thereof in anamount effective to increase muscle performance, muscle size and/ormuscle strength in a subject.

In certain embodiments, provided herein are methods of treating,preventing, suppressing, inhibiting or reducing the incidence of musclewasting in a subject. The methods include administering to the subject acompound of Formula I, II or III or pharmaceutically acceptable salts orprodrugs thereof in an amount effective to treat, prevent, suppress,inhibit or reduce the incidence of a muscle wasting in the subject. Insome embodiments, the muscle wasting is caused by a condition selectedfrom among andropause, spinal muscular atrophies, muscular dystrophies(e.g., Duchenne, Myotonic and Becker), myasthenia gravis, cachexias suchas AIDS cachexia, cardiac cachexia, and cancer cachexia, cancer, ChronicObstructive Pulmonary Disease (COPD), emphysema, diabetes, HIVinfection, acquired immunodeficiency syndrome (AIDS), sepsis,tuberculosis, renal failure, heart failure, cardiomyopathy, bed rest,disuse, inactivity, microgravity, malnutrition, sarcopenia, aging andfrailty (e.g., see Lynch et al., Pharmacology & Therapeutics 113(3):461-487 (2007)).

In certain embodiments, provided herein are methods of treating aneuro-degenerative disease or disorder in a subject. The methods includeadministering to the subject a compound of Formula I, II or III orpharmaceutically acceptable salts or prodrugs thereof in an amounteffective to treat the neurodegenerative disease or disorder in thesubject. In some embodiments, the neurodegenerative disorder isAlzheimer's disease.

In certain embodiments, provided herein are methods for preventing theonset or delaying the progression of Alzheimer's disease in a subject.The methods include administering to the subject a compound of FormulaI, II or III or pharmaceutically acceptable salts or prodrugs thereof inan amount effective to prevent the onset or delay the progression ofAlzheimer's disease in the subject. In some embodiments, the compound isco-administered with an effective amount of a compound that inhibits theformation or release of β-amyloid.

In certain embodiments, provided herein are methods for treatingcognitive impairment in a subject. The methods include administering tothe subject a compound of Formula I, II or III or pharmaceuticallyacceptable salts or prodrugs thereof in an amount effective to treatcognitive impairment in the subject.

In certain embodiments, provided herein are methods for treatingdepression in a subject. The methods include administering to thesubject a compound of Formula I, II or III or pharmaceuticallyacceptable salts or prodrugs thereof in an amount effective to treatdepression in the subject.

In certain embodiments, provided herein are methods for treating one ormore postmenopausal conditions in a subject. The methods includeadministering to the subject a compound of Formula I, II or III orpharmaceutically acceptable salts or prodrugs thereof in an amounteffective to treat one or more postmenopausal conditions in the subject.In some embodiments, the postmenopausal condition is selected from amongloss of libido, decreased sexual activity, diminished feelings ofphysical well-being, fatigue and hot flashes. In some embodiments, thecompound is co-administered with another therapeutic agent selected fromamong estrone, 2-hydroxyestrone, 2-methoxyestrone, 4-hydroxyestrone,15-α-hydroxy-estrone, 16-α-hydroxyestrone, 16-β-hydroxyestrone,estradiol (17-β-estradiol), 2-hydroxy-estradiol, 2-methoxy-estradiol,4-hydroxy-estradiol, 16-oxoestradiol, estriol, 16-epiestriol and17-epiestriol and combinations thereof. In some embodiments, thecompound is co-administered with another therapeutic agent selected fromamong estradiol valerate, estrone, estrone sulfate, an estrone sulfatepiperazine salt or an ester thereof, a synthetic estrogen andcombinations thereof. In some embodiments, the compound isco-administered with another therapeutic agent selected from amongalendronate, calcitonin, clodronate, clomiphene, clomiphene citrate,clonidine, conjugated estrogen, natural or synthetic estrogen, ethinylestradiol, estradiol, enclomiphene, enclomiphene citrate, etidronate,ibandronate, medroxy-progesterone acetate, megestrol acetate,norethindrone acetate, pamidronate, progesterone, risedronate,tiludronate, zuclomiphene, zuclomiphene citrate and combinationsthereof.

In certain embodiments, provided herein are methods of improving thelipid profile in a subject. The methods include administering to thesubject a compound of Formula I, II or III or pharmaceuticallyacceptable salts or prodrugs thereof, in an amount effective to affectthe lipid profile in the subject.

In certain embodiments, provided herein are methods of reducingcirculating lipid levels in a subject. The methods include administeringto the subject a compound of Formula I, II or III or pharmaceuticallyacceptable salts or prodrugs thereof, in an amount effective to reducecirculating lipid levels in the subject. In some embodiments, thecompound is co-administered with a therapeutic agent selected from amongβ-hydroxy-β-methylbutyric acid, lactoferrin, cholestyramine, colestipol,colesevelam, nicotinic acid, one or more fibric acids (e.g.,gemfibrozil, fenofibrate and clofibrate) and one or more HMG-CoAreductase inhibitors (lovastatin, pravastatin, simvastatin, fluvastatin,atorvastatin and cerivastatin) and combinations thereof.

In certain embodiments, provided herein are methods of treatingatherosclerosis, a cardiovascular disorder, a cerebrovascular disorder,a peripheral vascular disorder, and/or an intestinal vascular disorderin a subject. The methods include administering to the subject acompound of Formula I, II or III or pharmaceutically acceptable salts orprodrugs thereof, in an amount effective to treat the atherosclerosis,cardiovascular disorder, cerebrovascular disorder, peripheral vasculardisorder, and/or intestinal vascular disorder in the subject. In someembodiments, the compound is co-administered with a selective estrogenreceptor modulator (SERM) compound.

In certain embodiments, provided herein are methods of treatingosteoporosis, osteopenia, glucocorticoid-induced osteoporosis or bonefracture in a subject. The methods include administering to the subjecta compound of Formula I, II or III or pharmaceutically acceptable saltsor prodrugs thereof, in an amount effective to treat the osteoporosis,osteopenia, glucocorticoid-induced osteoporosis or bone fracture in thesubject. In some embodiments, the compound is co-administered with aneffective amount of at least one other therapeutic agent selected fromamong estrogen, estrogen derivatives, progestin, progestin derivatives,a bisphosphonate, an anti-estrogen, a selective estrogen receptormodulator (SERM), an α_(v)β₃ integrin receptor antagonist, a cathepsininhibitor, a proton pump inhibitor, a PPARγ inhibitor, calcitonin,osteoprotegerin and combinations thereof.

In certain embodiments, provided herein are methods of increasing thestrength or mass of bone of a subject. The methods include administeringto the subject a compound of Formula I, II or III or pharmaceuticallyacceptable salts or prodrugs thereof, in an amount effective to increasethe strength or mass of a bone in the subject.

In certain embodiments, provided herein are methods of promoting boneformation in a subject. The methods include administering to the subjecta compound of Formula I, II or III or pharmaceutically acceptable saltsor prodrugs thereof, in an amount effective to promote bone formation inthe subject.

In certain embodiments, provided herein are methods of treating ahematopoietic disorder in a subject. The methods include administeringto the subject a compound of Formula I, II or III or pharmaceuticallyacceptable salts or prodrugs thereof, in an amount effective to treatthe hematopoietic disorder in the subject. In some embodiments, thehematopoietic disorder is selected from among anemia, leukemia, andhematopoietic conditions caused by bone marrow transplantation orchemotherapy or radiation therapy.

In certain embodiments, provided herein are methods of increasing thenumber of red blood cells in a subject. The methods includeadministering to the subject a compound of Formula I, II or III orpharmaceutically acceptable salts or prodrugs thereof, in an amounteffective to increase the number of red blood cells in the subject.

In certain embodiments, provided herein are methods of treating anemia,thrombocytopenia or neutropenia in a subject. The methods includeadministering to the subject a compound of Formula I, II or III orpharmaceutically acceptable salts or prodrugs thereof, in an amounteffective to treat anemia, thrombocytopenia or neutropenia in thesubject. In some embodiments, the compound is co-administered with atherapeutically effective amount of at least one hematopoietic cytokine.In some embodiments, the hematopoietic cytokine is selected from amongerythropoietin, granulocyte-colony stimulating factor,granulocyte-macrophage-colony stimulating factor, interleukin-1,interleukin-3, interleukin-4, interleukin-5, interleukin-7,interleukin-9, interleukin-11, macrophage-colony stimulating factor,stem cell factor and thrombopoietin.

In certain embodiments, provided herein are methods of increasing serumerythropoietin (EPO) levels in a subject. The methods includeadministering to the subject a compound of Formula I, II or III orpharmaceutically acceptable salts or prodrugs thereof, in an amounteffective to increasing serum EPO levels in the subject.

In certain embodiments, provided herein are methods of preventing and/ortreating obesity or an obesity-related condition or disease in asubject. The methods include administering to the subject a compound ofFormula I, II or III or pharmaceutically acceptable salts or prodrugsthereof, in an amount effective to prevent and/or treat obesity or anobesity-related condition or disease in the subject.

In certain embodiments, provided herein are methods of treatingabdominal adiposity in a subject. The methods include administering tothe subject a compound of Formula I, II or III or pharmaceuticallyacceptable salts or prodrugs thereof that is an AR agonist, in an amounteffective to treat abdominal adiposity in the subject.

In certain embodiments, provided herein are methods of treatingabdominal obesity in a subject. The methods include administering to thesubject a compound of Formula I, II or III or pharmaceuticallyacceptable salts or prodrugs thereof that is an AR antagonist, in anamount effective to treat abdominal obesity in the subject.

In certain embodiments, provided herein are methods of treating insulinresistance in a subject. The methods include administering to thesubject a compound of Formula I, II or III or pharmaceuticallyacceptable salts or prodrugs thereof, in an amount effective to treatinsulin resistance in the subject.

In certain embodiments, provided herein are methods of treating type 2diabetes in a subject. The methods include administering to the subjecta compound of Formula I, II or III or pharmaceutically acceptable saltsor prodrugs thereof, in an amount effective to treat type 2 diabetes inthe subject. In some embodiments, the compound is co-administered withan effective amount of an anti-diabetic drug, such as, but not limitedto, thiazolidinedione-type drugs such as pioglitazone or rosiglitazone,sulfonylurea-type drugs, such as chlorpropamide, glimepiride, glipizide,glyburide or tolbutamide, a biguanide-type drug such as metformin,exenatide, acarbose, repaglinide, nateglinide, tolazamide orcombinations thereof.

In certain embodiments, provided herein are methods of treating arterialhypertension, hyper-insulinaemia, hyperglycemia or dyslipidemia in asubject. The methods include administering to the subject a compound ofFormula I, II or III or pharmaceutically acceptable salts or prodrugsthereof, in an amount effective to treat arterial hypertension,hyper-insulinaemia, hyperglycemia or dyslipidemia in the subject.

In certain embodiments, provided herein are methods for the treatment orprevention of an arthritic condition or inflammatory disorder in asubject. The methods include administering to the subject a compound ofFormula I, II or III or pharmaceutically acceptable salts or prodrugsthereof, in an amount effective to treat or prevent the arthriticcondition or inflammatory disorder in the subject. In some embodiments,the arthritic condition or inflammatory disorder is selected from amongosteoarthritis, Behcet's disease, bursitis, tendonitis, CPPD depositiondisease, carpal tunnel syndrome, Ehlers-Danlos syndrome, fibromyalgia,gout, infectious arthritis, inflammatory bowel disease, juvenilearthritis, lupus erythematosus, Lyme disease, Marfan syndrome, myositis,osteoarthritis, osteogenesis imperfecta, osteonecrosis, polyarteritis,polymyalgia rheumatica, psoriatic arthritis, Raynaud's phenomenon,reflex sympathetic dystrophy syndrome, Reiter's syndrome, rheumatoidarthritis, scleroderma and Sjogren's syndrome.

In certain embodiments, provided herein are methods for the treatment orprevention of osteoarthritis in a subject. The methods includeadministering to the subject a compound of Formula I, II or III orpharmaceutically acceptable salts or prodrugs thereof, in an amounteffective to treat or prevent osteoarthritis in the subject. In someembodiments, the compound is co-administered with corticosteroids, goldtreatment, methotrexate, aspirin, NSAIDs, COX-2 inhibitors and DMARDs(Disease-Modifying Anti-Rheumatic Drugs).

In certain embodiments, provided herein are methods of treating sexualdysfunction in a subject. The methods include administering to thesubject a compound of Formula I, II or III or pharmaceuticallyacceptable salts or prodrugs thereof, in an amount effective to treatsexual dysfunction in the subject. In some embodiments, the sexualdysfunction is male erectile dysfunction. In some embodiments, thesexual dysfunction is impotence.

In certain embodiments, provided herein are methods of increasing thelibido of a subject. The methods include administering to the subject acompound of Formula I, II or III or pharmaceutically acceptable salts orprodrugs thereof, in an amount effective to increase the libido of thesubject.

In certain embodiments, provided herein are methods of treating acondition related to androgen decline in a male subject. The methodsinclude administering to the subject a compound of Formula I, II or IIIor pharmaceutically acceptable salts or prodrugs thereof, in an amounteffective to treat the condition related to androgen decline in thesubject. In some embodiments, the condition is selected from amongfatigue, depression, decreased libido, sexual dysfunction, erectiledysfunction, hypogonadism, osteoporosis, hair loss, obesity, sarcopenia,osteopenia, benign prostate hyperplasia, anemia, alterations in mood andcognition, and prostate cancer. In certain embodiments, provided aremethods of treating sarcopenia in a subject.

In certain embodiments, provided herein are methods of treating acondition related to androgen deficiency in a female subject. Themethods include administering to the subject a compound of Formula I, IIor III or pharmaceutically acceptable salts or prodrugs thereof, in anamount effective to treat the condition related to androgen decline inthe subject. In some embodiments, the condition is selected from amongsexual dysfunction, decreased sexual libido, sarcopenia, osteopenia,osteoporosis, alterations in cognition and mood, depression, anemia,hair loss, obesity, endometriosis, breast cancer, uterine cancer andovarian cancer.

In certain embodiments, provided herein are methods of treating adisease in a subject. The methods include administering to the subject acompound of Formula I, II or III or pharmaceutically acceptable salts orprodrugs thereof, in an amount effective to treat the disease, whereinthe disease is selected from among angina, coronary artery disease,arteriosclerosis, atherosclerosis, obesity, diabetes, syndrome X,glucose intolerance, insulin resistance, hypercholesterolemia,hyperlipoproteinemia, hyper-glycemia, hyperinsulinemia, hyperlipidemia,glaucoma, hypertension, hypertriglyceridemia, renal disease, thrombosis,peripheral vascular disease, vascular wall damage, stroke, dyslipidemia,diabetic dyslipidemia, mixed dyslipidemia and nonalcoholic fatty liverdisease.

Pharmaceutical compositions formulated for administration by anappropriate route and means including effective concentrations of one ormore than one compound of Formula I, II or III or pharmaceuticallyacceptable salts or prodrugs thereof, or pharmaceutically acceptablederivatives thereof, that deliver amounts effective for the treatment,prevention, or amelioration of one or more symptoms of diseases ordisorders that are modulated or otherwise affected by androgen receptoractivity, or in which androgen receptor activity is implicated, also areprovided. The effective amounts and concentrations are effective forameliorating any of the symptoms of any of the diseases or disorders.

In certain embodiments, provided herein is a pharmaceutical compositionincluding: i) a physiologically acceptable carrier, diluent, and/orexcipient; and ii) one or more than one compound of Formula I, II or IIIor pharmaceutically acceptable salts or prodrugs thereof.

In certain embodiments, compounds provided herein are used to detect thepresence, quantity and/or state of androgen receptors in a sample, suchas a cell, cell homogenates and lysates. In some embodiments, samplesare obtained from a subject. In certain embodiments, compounds areradio- or isotopically-labeled.

Also provided are articles of manufacture that include packagingmaterial, within the packaging material one or more than one compound ofFormula I, II or III or pharmaceutically acceptable salts or prodrugsthereof, or composition that includes one or more than one compound ofFormula I, II or III or pharmaceutically acceptable salts or prodrugsthereof, that is effective for modulating the activity of androgenreceptor, or for treatment, prevention or amelioration of one or moresymptoms of androgen receptor mediated diseases or disorders, ordiseases or disorders in which androgen receptor activity is implicated,and a label that indicates that the compound or composition is used formodulating the activity of androgen receptor, or for treatment,prevention or amelioration of one or more symptoms of androgen receptormediated diseases or disorders, or diseases or disorders in whichandrogen receptor activity is implicated.

Provided herein also are kits, which contain the compositions includingthe compounds described herein, a device for administration of thecomposition and, optionally, instructions for administration.

DETAILED DESCRIPTION

A. Definitions

B. Compounds

C. Preparation of the Compounds

-   -   1. Scheme 1—Preparation of Compounds of Formula I    -   2. Scheme II and III—Preparation of Compounds of Formula II    -   3. Scheme IV—Preparation of Compounds of Formula III

D. Certain Indications

-   -   1. Muscle Wasting    -   2. Muscle Tone and Strength    -   3. Osteoporosis    -   4. Prostate Disease and Prostate Cancer    -   5. Hematopoietic Conditions and Disorders    -   6. Neurodegenerative Diseases and Disorders    -   7. Obesity    -   8. Insulin Disorders and Diabetes    -   9. Sexual Dysfunction    -   10. Arthritic Conditions and Inflammatory Disorders    -   11. Modifying Lipid Profile    -   12. Contraception    -   13. Postmenopausal Conditions

E. Formulation of Pharmaceutical Compositions

-   -   1. Compositions for Oral Administration    -   2. Injectables, Solutions and Emulsions    -   3. Lyophilized Powders    -   4. Topical Administration    -   5. Compositions for Other Routes of Administration

F. Articles of Manufacture

G. Kits

H. Evaluation of the Activity of the Compounds

-   -   1. Effect on muscle    -   2. Effect on bone    -   3. Antagonist activity against hormone-dependent tumors    -   4. Efficacy and toxicity    -   5. Receptor Binding Assays    -   6. In vivo assay—Sprague-Dawley Rat Models

I. Methods of Use of the Compounds and Compositions

-   -   1. Methods of Treating Muscle Wasting    -   2. Methods for Improving Muscle Performance, Size and/or        Strength    -   3. Methods of Improving Athletic Performance    -   4. Methods of Treating Bone-related conditions    -   5. Methods of Treating Cancer    -   6. Methods of Treating Prostate Cancer    -   7. Methods of Contraception    -   8. Methods of Providing Hormone Therapy    -   9. Methods of Treating Postmenopausal Conditions    -   10. Methods of Treating Hematopoietic Disorders    -   11. Methods of Treating Neurodegenerative Diseases and Disorders    -   12. Methods of Treating Cognitive Impairment    -   13. Methods of Treating Depression    -   14. Methods of Treating Obesity    -   15. Methods of Treating Insulin Resistance and Diabetes    -   16. Methods of Treating Sexual Dysfunction    -   17. Methods of Treating Arthritic Conditions and Inflammatory        Disorders    -   18. Methods of Improving Lipid Profile    -   19. Methods of Treating Atherosclerosis    -   20. Methods of Treating Conditions Related to Androgen Decline    -   21. Methods of Treating Conditions Related to Androgen        Deficiency

J. Combination Therapies

K. Examples

The section headings used herein are for organizational purposes onlyand are not to be construed as limiting the subject matter described.All documents, or portions of documents, cited in the applicationincluding, but not limited to, patents, patent applications, articles,books, manuals, and treatises are hereby expressly incorporated byreference in their entirety for any purpose.

Unless specific definitions are provided, the nomenclatures used inconnection with, and the laboratory procedures and techniques of,analytical chemistry, synthetic organic chemistry, and medicinal andpharmaceutical chemistry described herein are those known in the art.Standard techniques can be used for chemical syntheses, chemicalanalyses, pharmaceutical preparation, formulation, and delivery, andtreatment of subjects. Reactions and purification techniques can beperformed e.g., using kits according to manufacturer's specifications oras commonly accomplished in the art or as described herein. Theforegoing techniques and procedures can be generally performed accordingto conventional methods well known in the art and as described invarious general and more specific references that are cited anddiscussed herein. See e.g., Sambrook et al. Molecular Cloning: ALaboratory Manual (2d ed., Cold Spring Harbor Laboratory Press, ColdSpring Harbor, N.Y. (1989)), which is incorporated herein by referencefor any purpose.

A. DEFINITIONS

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as is commonly understood by one of skill in theart to which the claimed subject matter belongs. All patents, patentapplications, published materials referred to throughout the entiredisclosure herein, unless noted otherwise, are incorporated by referencein their entirety. In the event that there are a plurality ofdefinitions for terms herein, those in this section prevail. Wherereference is made to a URL or other such identifier or address, itunderstood that such identifiers can change and particular informationon the internet can come and go, but equivalent information can be foundby searching the internet. Reference thereto evidences the availabilityand public dissemination of such information.

It is to be understood that both the foregoing general description andthe following detailed description are exemplary and explanatory onlyand are not restrictive of the subject matter claimed. In thisapplication, the use of the singular includes the plural unlessspecifically stated otherwise. In this application, the use of “or”means “and/or” unless stated otherwise. Furthermore, use of the term“including” as well as other forms, such as “includes,” and “included”is not limiting.

As used herein, ranges and amounts can be expressed as “about” aparticular value or range. “About” also includes the exact amount. Hence“about 10%” means “about 10%” and also “10%.”

As used herein, “optional” or “optionally” means that the subsequentlydescribed event or circumstance does or does not occur, and that thedescription includes instances where the event or circumstance occursand instances where it does not. For example, an optionally substitutedgroup means that the group is unsubstituted or is substituted.

As used herein, the singular forms “a,” “an” and “the” include pluralreferents unless the context clearly dictates otherwise. Thus, forexample, reference to a composition comprising “a therapeutic agent”includes compositions with one or a plurality of therapeutic agents.

As used herein, the term “target receptor” refers to a molecule or aportion of a receptor capable of being bound by a selective bindingcompound. In certain embodiments, a target receptor is a androgenreceptor.

As used herein, the term “selective binding compound” refers to acompound that selectively binds to any portion of one or more targetreceptors.

As used herein, the term “selectively binds” refers to the ability of aselective binding compound to bind to a target receptor with greateraffinity than it binds to a non-target receptor. In certain embodiments,specific binding refers to binding to a target with an affinity that isat least 2, 5, 10, 25, 50, 75, 100, 150, 200, 250, 500, 1000 or moretimes greater than the affinity for a non-target.

As used herein, the term “androgen receptor selective binding compound”refers to a compound that selectively interacts with an androgenreceptor with a greater affinity than it with a non-androgen receptor,such as, but not limited to, a progesterone receptor (PR), estrogenreceptor (ER), glucocorticoid receptor (GR), mineralocorticoid receptor(MR), retinoic acid receptor (RAR), rexinoid receptor (RXR), orperoxisome proliferator-activated receptor (PPAR). In certainembodiments, an androgen receptor selective binding compound binds to anandrogen receptor with an affinity that is at least 5, 10, 25, 50, 75,100, 150, 200, 250, 500, 1000 or more times greater than the affinityfor a non-androgen receptor. In some embodiments, the compounds providedherein are androgen receptor selective binding compounds.

As used herein, “treating a subject having a disease or condition” meansthat a compound, composition or other product provided herein isadministered to the subject.

As used herein, the terms “treat” and “treating” encompass either orboth responsive and prophylaxis measures, e.g., designed to inhibit,slow or delay the onset of a symptom of a disease or disorder, achieve afull or partial reduction of a symptom or disease state, and/or toalleviate, ameliorate, lessen, or cure a disease or disorder and/or itssymptoms. The term treatment also is intended to include prophylactictreatment.

As used herein, the term “treatment” means any manner in which thesymptoms of a condition, disorder or disease are ameliorated orotherwise beneficially altered. Hence, treatment encompassesprophylaxis, therapy and/or cure. Treatment also encompasses anypharmaceutical use of the compositions herein.

As used herein, the term “therapeutic agent” refers to conventionaldrugs and drug therapies, including vaccines, which are known to thoseskilled in the art.

As used herein, amelioration of the symptoms of a particular disease ordisorder by a treatment, such as by administration of a compoundprovided herein or a pharmaceutical composition thereof or othertherapeutic agent, refers to any lessening, whether permanent ortemporary, lasting or transient, of the symptoms that can be attributedto or associated with administration of the composition or therapeutic.

As used herein, prevention or prophylaxis refers to methods in which therisk of developing disease or condition is reduced. Prophylaxis includesreduction in the risk of developing a disease or condition and/or aprevention of worsening of symptoms or progression of a disease orreduction in the risk of worsening of symptoms or progression of adisease.

As used herein, an effective amount of a compound or composition fortreating a particular disease is an amount that is sufficient toameliorate, or in some manner reduce the symptoms associated with thedisease. Such amount can be administered as a single dosage or can beadministered according to a regimen, whereby it is effective. The amountcan cure the disease but, typically, is administered in order toameliorate the symptoms of the disease. Typically, repeatedadministration is required to achieve a desired amelioration ofsymptoms.

As used herein, “therapeutically effective amount” or “therapeuticallyeffective dose” refers to an agent, compound, material, or compositioncontaining a compound that is at least sufficient to produce atherapeutic effect. An effective amount is the quantity of a therapeuticagent necessary for preventing, curing, ameliorating, arresting orpartially arresting a symptom of a disease or disorder.

As used herein, amelioration of the symptoms of a particular disorder byadministration of a particular compound or pharmaceutical compositionrefers to any lessening of severity, delay in onset, slowing ofprogression, or shortening of duration, whether permanent or temporary,lasting or transient, that can be attributed to or associated withadministration of the compound or composition.

As used herein, the term “modulator” refers to a compound that alters anactivity of a molecule. For example, a modulator can cause an increaseor decrease in the magnitude of a certain activity of a moleculecompared to the magnitude of the activity in the absence of themodulator. In certain embodiments, a modulator is an inhibitor, whichdecreases the magnitude of one or more activities of a molecule. Incertain embodiments, an inhibitor completely prevents one or moreactivities of a molecule. In certain embodiments, a modulator is anactivator, which increases the magnitude of at least one activity of amolecule. In certain embodiments, the presence of a modulator results inan activity that does not occur in the absence of the modulator.

As used herein, the term “selective modulator” refers to a compound thatselectively modulates a target activity.

As used herein, “selective androgen receptor modulator” or “SARM” is acompound that mimics the action of a natural androgen receptor ligand insome tissues but not in others. SARMs are compounds that elicit androgenagonism in one or more target tissues (e.g., muscle and/or bone) andantagonism and/or minimal agonism or no effect in other tissues (e.g.,skin, prostate). SARMs exhibit tissue selective androgen agonism. Amongthe compounds provided herein, are those that are SARMs that exhibitagonistic anabolic properties and antagonistic androgenic properties inselected tissues. Others of the compounds are SARMs that are AR agonistsin some tissues and cause increased transcription of AR-responsive genes(e.g., muscle anabolic effect). In other tissues, compounds arecompetitive inhibitors of androgens such as testosterone on the AR andthereby prevent agonistic effects of the native androgens. For example,compounds provided herein are SARMs that have agonist activity in muscleand demonstrate antagonist activity in a gonad of a subject. SARMs thatdemonstrate such activity can increase muscle mass and decrease fat insubjects without causing androgenic side effects, such as sebaceousgland stimulation.

As used herein, “tissue selective androgen receptor agonism” refers tothe ability of a SARMs compound to agonize an androgen receptor of one(or more than one) target tissue with greater affinity than it agonizesan androgen receptor of a non-target tissue.

In certain embodiments, tissue selective androgen receptor agonismrefers to agonism of an AR of a target tissue that is at least about or2 fold up to more than about or 500 fold, greater than the androgenreceptor agonism of an androgen receptor of a non-target tissue.

In certain embodiments, tissue selective androgen receptor agonismrefers to agonism of an AR of a target tissue that is at least 2, 5, 10,15, 20, 25, 30, 50, 60, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300,350, 400, 450, 500, 600, 700, 800, 900 or 1000 or more times greaterthan the androgen receptor agonism of an androgen receptor of anon-target tissue. For example, SARMs can exhibit agonism of an ARreceptor in muscle tissue and antagonism of the AR in prostate tissue.

As used herein, “tissue selective androgen receptor antagonism” refersto the ability of a SARMs compound to antagonize an androgen receptor ofone (or more than one) target tissue with greater affinity than itantagonizes an androgen receptor of a non-target tissue. In certainembodiments, tissue selective androgen receptor antagonism refers toantagonism of an AR of a target tissue that is at least about or 2 foldup to more than about or 500 fold, greater than the androgen receptoragonism of an androgen receptor of a non-target tissue. In certainembodiments, tissue selective androgen receptor antagonism refers toantagonism of an AR of a target tissue that is at least 2, 5, 10, 15,20, 25, 30, 50, 60, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300,350, 400, 450, 500, 600, 700, 800, 900 or 1000 or more times greaterthan the androgen receptor antagonism of an androgen receptor of anon-target tissue. For example, SARMs can exhibit antagonist activityagainst hormone-dependent tumors while exhibiting no activity, or insome instances agonist activity, against other non-tumor tissuescontaining the androgen receptor.

As used herein, the term “selectively modulates” refers to the abilityof a selective modulator to modulate a target activity to a greaterextent than it modulates a non-target activity. In certain embodimentsthe target activity is selectively modulated by, for example about or 2fold up to more than about or 500 fold, in some embodiments, about or 2,5, 10, 25, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450 or more than500 fold.

As used herein, an “activity” of a SARMS compound provided herein refersto any activity exhibited by a selective androgen modulator. Suchactivities can be tested in vitro and/or in vivo and include, but arenot limited to, agonism or antagonism of an androgen receptor. Activitycan be assessed in vitro or in vivo using recognized assays, forexample, by using the co-transfection assay. The results of such assaysthat indicate that a compound exhibits an activity can be correlated toactivity of the compound in vivo, in which in vivo activity can bereferred to as biological activity. Assays to determine functionality oractivity of androgen receptor modulators, including selective androgenreceptor modulator compounds, are known to those of skill in the art.Exemplary assays include, but are not limited to, fluorescencepolarization assay, luciferase assay and co-transfection assay. Incertain embodiments, the compounds provided herein are capable ofmodulating activity of androgen receptor in a “co-transfection” assay(also called a “cis-trans” assay), which is known in the art (see e.g.,Evans et al., Science 240: 889-895 (1988); U.S. Pat. Nos. 4,981,784 and5,071,773; and Pathirana et al., “Nonsteroidal Human ProgesteroneReceptor Modulators from the Marie Alga Cymopolia Barbata,” Mol. Pharm.47: 630-35 (1995)).

As used herein, “biological activity” refers to the in vivo activitiesof a compound or physiological responses that result upon in vivoadministration of a compound, composition or other mixture. Biologicalactivity, thus, encompasses therapeutic effects and pharmaceuticalactivity of such compounds, compositions and mixtures. Biologicalactivities can be observed in in vitro systems designed to test or usesuch activities. Thus, for purposes herein a biological activity of aselective androgen receptor modulator encompasses the agonism orantagonism of an androgen receptor.

As used herein, the term “assess” and grammatical variations thereof, isintended to include quantitative and qualitative determination in thesense of obtaining an absolute value for the activity of a compound, andalso of obtaining an index, ratio, percentage, visual or other valueindicative of the level of the activity. Assessment can be direct orindirect.

As used herein, the term “targeting agent” refers to any moiety, such asa protein or effective portion thereof, that provides specific bindingto a cell surface molecule, such a cell surface receptor, which in someinstances can internalize a bound conjugate or portion thereof. Atargeting agent also can be one that promotes or facilitates, forexample, affinity isolation or purification of the conjugate; attachmentof the conjugate to a surface; or detection of the conjugate orcomplexes containing the conjugate.

As used herein, derivative or analog of a molecule refers to a portionderived from or a modified version of the molecule. In some embodiments,a derivative includes, but is not limited, to acid derivatives, amidederivatives, ester derivatives and ether derivatives. In otherembodiment, the SARM compounds provided herein are hydrates, includinghemihydrate, monohydrate, dehydrate and trihydrate.

As used herein, the term “disease” or “disorder” refers to apathological condition in an organism resulting from cause or conditionincluding, but not limited to, infections, acquired conditions, geneticconditions, and characterized by identifiable symptoms. Diseases anddisorders also include those that are caused by the absence of acompound, such as an androgen agonist.

As used herein, “patient” or “subject” to be treated includes humans andor non-human animals, including mammals. Mammals include primates, suchas humans, chimpanzees, gorillas and monkeys; domesticated animals, suchas dogs, horses, cats, pigs, goats, cows; and rodents such as mice,rats, hamsters and gerbils.

As used herein, “animal” includes any animal, such as, but not limitedto; primates including humans, gorillas and monkeys; rodents, such asmice and rats; fowl, such as chickens; ruminants, such as goats, cows,deer, sheep; ovine, such as pigs and other animals. Non-human animalsexclude humans as the contemplated animal.

As used herein, a “combination” refers to any association between two oramong more items. The association can be spatial or refer to the use ofthe two or more items for a common purpose.

As used herein, a “composition” refers to any mixture of two or moreproducts or compounds (e.g., agents, modulators, regulators, etc.). Itcan be a solution, a suspension, liquid, powder, a paste, aqueous ornon-aqueous formulations or any combination thereof.

As used herein, a “fluid” refers to any composition that can flow.Fluids thus encompass compositions that are in the form of semi-solids,pastes, solutions, aqueous mixtures, gels, lotions, creams and othersuch compositions.

As used herein, a “kit” refers to a packaged combination, optionallyincluding reagents and other products and/or components for practicingmethods using the elements of the combination. For example, kitscontaining a compound provided herein and another item for a purposeincluding, but not limited to, administration, diagnosis, and assessmentof a biological activity or property are provided. Kits optionallyinclude instructions for use.

As used herein, an “article of manufacture” is a product that is madeand sold. As used throughout this application, the term is intended toencompass compounds of formulae I, II or III contained in articles ofpackaging.

As used herein, the term “target activity” refers to a target activitycapable of being modulated by a selective modulator. Certain exemplarytarget activities include, but are not limited to, binding affinity,signal transduction, enzymatic activity, tumor growth, inflammation orinflammation-related processes, and amelioration of one or more symptomsassociated with a disease or condition.

As used herein, the term “mediate” means affect or influence. Thus, forexample, conditions mediated by an androgen receptor are those in whichan androgen receptor plays a role. Androgen receptors are known to playa role in conditions including, for example, acne, aging skin,male-pattern baldness, sexual dysfunction, impotence, depression,wasting diseases, HIV-wasting, frailty, cognitive decline, Alzheimer'sdisease, sleep apnea, hirsutism, hypogonadism, premature ovarianfailure, inflammatory arthritis and joint repair, osteopenia,osteoporosis, glucocorticoid-induced osteoporosis, bone fracture, bonedamage following bone reconstructive surgery, atherosclerosis,hypercholesterolemia, hyperlipidemia, aplastic anemia and otherhematopoietic disorders, obesity, abdominal adiposity, metabolicsyndrome, type II diabetes, muscular dystrophies, periodontal disease,sarcopenia, postmenopausal symptoms in women, prostatic hyperplasia,prostate cancer, benign prostatic hyperplasia (BPH), cancer cachexia,and hormone-dependent cancers.

As used herein, the term “receptor mediated activity” refers anybiological activity that results, either directly or indirectly, frombinding of a ligand to a receptor.

As used herein, the term “agonist” refers to a compound, the presence ofwhich results in an activity of a receptor that is the same as theactivity resulting from the presence of a naturally occurring ligand forthe receptor. An agonist of the androgen receptor can bind to theandrogen receptor and initiate a physiological or a pharmacologicalresponse characteristic of that receptor. A “full agonist” induces fullactivation of the androgen receptor population at a given concentration.

As used herein, the term “partial agonist” refers to a compound thepresence of which results in a biological activity of a receptor that isof the same type as that resulting from the presence of a naturallyoccurring ligand for the receptor, but of a lower magnitude. A “partialagonist” is an agonist that is unable to induce maximal activation ofthe receptor population, regardless of the amount of compound applied.

As used herein, the term “antagonist” refers to a compound, the presenceof which results in a decrease in the magnitude of an activity of areceptor. In certain embodiments, the presence of an antagonist resultsin complete inhibition of an activity of a receptor. In anotherembodiment, the compound binds to androgen receptor and blocks orinhibits the androgen-associated responses normally induced by a naturalandrogen receptor ligand.

As used herein, the IC₅₀ refers to an amount, concentration or dosage ofa particular test compound that achieves a 50% inhibition of a maximalresponse, such as modulation of androgen receptor activity, in an assaythat measures such response. IC₅₀ also refers to the concentration oftest compound required to decrease specific binding by 50%. IC₅₀ valuescan be determined using the log-logit (Hill) method.

As used herein, EC₅₀ refers to a dosage, concentration or amount of aparticular test compound that elicits a dose-dependent response at 50%of maximal expression of a particular response that is induced, provokedor potentiated by the particular test compound.

As used herein, K_(i) values can be determined using the Cheng-Prusoffequation using a previously determined K_(d) value for the steroid, suchas dihydrotestosterone:K _(i)=IC₅₀/(1+[L]/K _(d))where [L]=concentration of labeled steroid and K_(d)=dissociationconstant of labeled steroid. For a discussion of the calculation ofK_(i), see e.g., Cheng, Y. C. and Prusoff, W. H. Biochem. Pharmacol. 22:3099 (1973).

As used herein, the term “carrier” refers to a compound that facilitatesthe incorporation of another compound into cells or tissues. Forexample, dimethyl sulfoxide (DMSO) is a carrier commonly used forimproving incorporation of certain organic compounds into cells ortissues.

As used herein, the term “pharmaceutical composition” refers to achemical compound or composition capable of inducing a desiredtherapeutic effect in a subject. In certain embodiments, apharmaceutical composition includes an active agent, which is the agentthat induces the desired therapeutic effect. In certain embodiments, apharmaceutical composition includes a prodrug. In certain embodiments, apharmaceutical composition includes inactive ingredients such ascarriers and excipients.

As used herein, a “prodrug” refers to a compound that is converted froma less active form into a corresponding more active form in vivo. Incertain embodiments, upon in vivo administration, a prodrug ischemically converted to the biologically, pharmaceutically ortherapeutically more active form of the compound. In certainembodiments, a prodrug is enzymatically metabolized by one or more stepsor processes to the biologically, pharmaceutically or therapeuticallyactive form of the compound. To produce a prodrug, a pharmaceuticallyactive compound is modified such that the active compound will beregenerated upon in vivo administration. The prodrug can be designed toalter the metabolic stability or the transport characteristics of adrug, to mask side effects or toxicity, to improve the flavor of a drugor to alter other characteristics or properties of a drug. By virtue ofknowledge of pharmacodynamic processes and drug metabolism in vivo,those of skill in this art, once a pharmaceutically active compound isknown, can design prodrugs of the compound (see, e.g., Nogrady,Medicinal Chemistry A Biochemical Approach, Oxford University Press, NewYork, pages 388-392 (1985)). Conventional procedures for the selectionand preparation of suitable prodrug derivatives are described, forexample, in Design of Prodrugs, (ed. H. Bundgaard, Elsevier, 1985),which is hereby incorporated herein by reference in its entirety. Anon-limiting example of a prodrug for use herein includes those thatpromote the solubility of alcohols such as by the procedures describedin Mahfous, N. H. et al, J. Pharm. Pharmacol. 53: 841-848 (2001) andBundgaard, H. et al., J. Med. Chem. 32: 2503-2507 (1989), both of whichare incorporated herein by reference in their entirety. Prodrugs includecompounds where hydroxy, ester, amine or sulfhydryl groups are bonded toany group that, when administered to a mammalian subject, cleaves toform a free hydroxyl, ester, amino, or sulfhydryl group, respectively.Examples of prodrugs include, but are not limited to, acetate, formateand benzoate derivatives of alcohol functional groups within thecompounds provided herein.

As used herein, the term “ester” refers to a chemical moiety withformula —(R)_(n)—COOR′, where R and R′ are independently selected fromthe group consisting of alkyl, cycloalkyl, aryl, heteroaryl (bondedthrough a ring carbon), a non-aromatic heterocycle, arylalkyl orheteroarylalkyl, and where n is 0 or 1.

Any hydroxy side chain on the compounds described herein can beesterified. The procedures and specific groups to be used to achievethis end are known to those of skill in the art and can readily be foundin reference sources such as Greene and Wuts, Protective Groups inOrganic Synthesis (3^(rd) ed., John Wiley & Sons, New York, N.Y.(1999)).

An example of a prodrug is a “prodrug ester” or “ester derivative” ofthe compounds disclosed herein, which are formed by the addition of anyof several ester-forming groups that are hydrolyzed under physiologicalconditions. Examples of prodrug ester groups include pivoyloxymethyl,acetoxymethyl, phthalidyl, indanyl and methoxymethyl, as well as othersuch groups known in the art. Other examples of prodrug ester groups canbe found in, for example, T. Higuchi and V. Stella, in “Prodrugs asNovel Delivery Systems”, Vol. 14, A.C.S. Symposium Series, AmericanChemical Society (1975); and “Bioreversible Carriers in Drug Design:Theory and Application”, edited by E. B. Roche, Pergamon Press: NewYork, 14-21 (1987).

As used herein, the term “pharmaceutically acceptable formulation”refers to a formulation of a compound that does not significantlyabrogate the biological activity, a pharmacological activity and/orother properties of the compound when the formulated compound isadministered to a subject. In certain embodiments, a pharmaceuticallyacceptable formulation does not cause significant irritation to asubject.

As used herein, “pharmaceutically acceptable derivative” refers toderivatives of a compound that does not significantly abrogate thebiological activity, a pharmacological activity and/or other propertiesof the compound when the formulated compound is administered to asubject, and include, but are not limited to, salts, esters, enolethers, enol esters, acetals, ketals, orthoesters, hemiacetals,hemiketals, acids, bases, solvates, hydrates or prodrugs thereof. Suchderivatives can be readily prepared by those of skill in this art usingknown methods for such derivatization. The compounds produced can beadministered to animals or humans without substantial toxic effects andeither are pharmaceutically active or are prodrugs.

As used herein, the term “pharmaceutically acceptable salt” is intendedto include all salts known and used in the art of pharmaceuticals.Pharmaceutically acceptable salts include, but are not limited to, aminesalts, such as but not limited to chloroprocaine, choline,N,N′-dibenzyl-ethylenediamine, ammonia, diethanolamine and otherhydroxyalkylamines, ethylenediamine, N-methylglucamine, procaine,N-benzyl-phenethylamine,1-para-chloro-benzyl-2-pyrrolidin-1′-ylmethyl-benzimidazole,diethylamine and other alkylamines, piperazine andtris(hydroxy-methyl)aminomethane; alkali metal salts, such as but notlimited to lithium, potassium and sodium; alkali earth metal salts, suchas but not limited to barium, calcium and magnesium; transition metalsalts, such as but not limited to zinc; and other metal salts, such asbut not limited to sodium hydrogen phosphate and disodium phosphate; andalso including, but not limited to, salts of mineral acids, such as butnot limited to hydrochlorides and sulfates; and salts of organic acids,such as but not limited to acetates, lactates, malates, tartrates,citrates, ascorbates, succinates, butyrates, valerates and fumarates.Exemplary pharmaceutically acceptable salts include acetate,lactobionate, benzenesulfonate, laurate, benzoate, malate, bicarbonate,maleate, bisulfate, mandelate, bitartrate, mesylate, borate,methylbromide, methylnitrate, calcium edetate, methylsulfate, camsylate,mucate, carbonate, napsylate, bromide, chloride, nitrate, clavulanate,N-methylglucamine, citrate, ammonium salt, dihydrochloride, oleate,edetate, oxalate, edisylate, pamoate (embonate), estolate, palmitate,esylate, pantothenate, fumarate, phosphate/diphosphate, gluceptate,polygalacturonate, gluconate, salicylate, glutamate, stearate,glycollylarsanilate, sulfate, hexylresorcinate, subacetate, hydrabamine,succinate, hydrobromide, tannate, hydrochloride, tartrate,hydroxynaphthoate, teoclate, iodide, tosylate, isothionate,triethiodide, lactate, panoate and valerate, which can be used as adosage form for modifying the solubility or hydrolysis characteristicsor can be used in sustained release or pro-drug formulations. Thepreparation of the pharmaceutically acceptable salts described above andother typical pharmaceutically acceptable salts is more fully describedby Berg et al., “Pharmaceutical Salts,” J. Pharm. Sci. 66: 1-19 (1977).

Pharmaceutically acceptable esters include, but are not limited to,alkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl, heteroaralkyl,cycloalkyl and heterocyclyl esters of acidic groups, including, but notlimited to, carboxylic acids, phosphoric acids, phosphinic acids,sulfonic acids, sulfinic acids and boronic acids. Pharmaceuticallyacceptable enol ethers include, but are not limited to, derivatives offormula C═C(OR) where R is hydrogen, alkyl, alkenyl, alkynyl, aryl,heteroaryl, aralkyl, heteroaralkyl, cycloalkyl or heterocyclyl.Pharmaceutically acceptable enol esters include, but are not limited to,derivatives of formula C═C(OC(O)R) where R is hydrogen, alkyl, alkenyl,alkynyl, aryl, heteroaryl, aralkyl, heteroaralkyl, cycloalkyl orheterocyclyl. Pharmaceutically acceptable solvates and hydrates arecomplexes of a compound with one or more solvent or water molecules, or1 to about or 100, or 1 to about or 10, or one to about or 2, 3 or 4,solvent or water molecules.

As used herein, the term “alkyl” refers to straight or branched chainsubstituted or unsubstituted hydrocarbon groups, in one embodiment 1 to40 carbon atoms, in another embodiment, 1 to 20 carbon atoms, in anotherembodiment, 1 to 10 carbon atoms. The expression “lower alkyl” refers toan alkyl group of 1 to 6 carbon atoms. An alkyl group can be a“saturated alkyl,” meaning that it does not contain any alkene or alkynegroups and in certain embodiments, alkyl groups are optionallysubstituted. An alkyl group can be an “unsaturated alkyl,” meaning thatit contains at least one alkene or alkyne group. An alkyl group thatincludes at least one carbon-carbon double bond (C═C) also is referredto by the term “alkenyl,” and in certain embodiments, alkenyl groups areoptionally substituted. An alkyl group that includes at least onecarbon-carbon triple bond (C≡C) also is referred to by the term“allynyl,” and in certain embodiments, allynyl groups are optionallysubstituted.

In certain embodiments, an alkyl contains 1 to 20 carbon atoms (wheneverit appears herein, a numerical range such as “1 to 20” refers to eachinteger in the given range; e.g., “1 to 20 carbon atoms” means that analkyl group can contain only 1 carbon atom, 2 carbon atoms, 3 carbonatoms, etc., up to and including 20 carbon atoms, although the term“alkyl” also includes instances where no numerical range of carbon atomsis designated). An alkyl can be designated as “C₁-C₄ alkyl” or bysimilar designations. By way of example only, “C₁-C₄ alkyl” indicates analkyl having one, two, three, or four carbon atoms, i.e., the alkyl isselected from among methyl, ethyl, propyl, iso-propyl, n-butyl,iso-butyl, sec-butyl and t-butyl. Thus “C₁-C₄” includes C₁-C₂, C₁-C₃,C₂-C₃ and C₂-C₄ alkyl. Alkyls include, but are not limited to, methyl,ethyl, propyl, isopropyl, butyl, isobutyl, tertiary butyl, pentyl,hexyl, ethenyl, propenyl, butenyl, hexenyl, ethynyl, propynyl, butynyland hexynyl.

As used herein, the term “haloalkyl” alone or in combination refers toan alkyl in which at least one hydrogen atom is replaced with a halogenatom. In certain of the embodiments in which two or more hydrogen atomare replaced with halogen atoms, the halogen atoms are all the same asone another. In certain of such embodiments, the halogen atoms are notall the same as one another. Certain haloalkyls are saturatedhaloalkyls, which do not include any carbon-carbon double bonds or anycarbon-carbon triple bonds. Certain haloalkyls are haloalkenes, whichinclude one or more carbon-carbon double bonds. Certain haloalkyls arehaloalkynes, which include one or more carbon-carbon triple bonds. Incertain embodiments, haloalkyls are optionally substituted.

Where the number of any given substituent is not specified (e.g.,“haloalkyl”), there may be one or more substituents present. Forexample, “haloalkyl” can include one or more of the same or differenthalogens. For example, “haloalkyl” includes each of the substituentsCF₃, CHF₂ and CH₂F.

As used herein, “pseudohalogen” refers to compounds that behavesubstantially similar to halides/halogens. Such compounds can be used inthe same manner and treated in the same manner as halides/halogens (X—,in which X is a halogen, such as Cl, F or Br). Pseudohalogens include,but are not limited to, cyanide, cyanate, thiocyanate, selenocyanate,trifluoromethoxy, trifluoromethyl and azide.

As used herein, “cycloalkyl” refers to a saturated mono- or multicyclicring system where each of the atoms forming a ring is a carbon atom.Cycloalkyls can be formed by three, four, five, six, seven, eight, nine,or more than nine carbon atoms. In one embodiment, the ring systemincludes 3 to 12 carbon atoms. In another embodiment, they ring systemincludes 3 to 6 carbon atoms. The term “cycloalkyl” includes rings thatcontain one or more unsaturated bonds. As used herein, the terms“cycloalkenyl” and “cycloalkynyl” are unsaturated cycloalkyl ringsystem. Cycloalkyls can be optionally substituted. In certainembodiments, a cycloalkyl contains one or more unsaturated bonds.Examples of cycloalkyls include, but are not limited to, cyclopropane,cyclobutane, cyclopentane, cyclopentene, cyclopentadiene, cyclohexane,cyclohexene, 1,3-cyclohexadiene, 1,4-cyclohexadiene, cycloheptane andcycloheptene.

As used herein, the term “aryl” refers to a monocyclic, bicyclic ortricyclic aromatic system that contains no ring heteroatoms. Where thesystems are not monocyclic, the term aryl includes for each additionalring the saturated form (perhydro form) or the partially unsaturatedform (for example the dihydro form or tetrahydro form) or the maximallyunsaturated (nonaromatic) form. In some embodiments, the term arylrefers to bicyclic radicals in which the two rings are aromatic andbicyclic radicals in which only one ring is aromatic. Examples of arylinclude phenyl, naphthyl, anthracyl, indanyl, 1,2-dihydro-naphthyl,1,4-dihydronaphthyl, indenyl, 1,4-naphthoquinonyl and1,2,3,4-tetrahydronaphthyl.

Aryl rings can be formed by three, four, five, six, seven, eight, nine,or more than nine carbon atoms. In some embodiments, aryl refers to a3-, 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11-, 12-, 13- or 14-membered, aromaticmono-, bi- or tricyclic system. In some embodiments, aryl refers to anaromatic C₃-C₉ ring. In some embodiments, aryl refers to an aromaticC₄-C₈ ring. Aryl groups can be optionally substituted.

As used herein, the term “heteroaryl” refers to an aromatic ring inwhich at least one atom forming the aromatic ring is a heteroatom.Heteroaryl rings can be formed by three, four, five, six, seven, eight,nine and more than nine atoms. Heteroaryl groups can be optionallysubstituted. Examples of heteroaryl groups include, but are not limitedto, aromatic C₃₋₈ heterocyclic groups containing one oxygen or sulfuratom, or two oxygen atoms, or two sulfur atoms or up to four nitrogenatoms, or a combination of one oxygen or sulfur atom and up to twonitrogen atoms, and their substituted as well as benzo- and pyrido-fusedderivatives, for example, connected via one of the ring-forming carbonatoms. In certain embodiments, heteroaryl is selected from amongoxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, pyridinyl,pyridazinyl, pyrimidinal, pyrazinyl, indolyl, benzimidazolyl,quinolinyl, isoquinolinyl, quinazolinyl or quinoxalinyl.

In some embodiments, a heteroaryl group is selected from among pyrrolyl,furanyl (furyl), thiophenyl (thienyl), imidazolyl, pyrazolyl,1,2,3-triazolyl, 1,2,4-triazolyl, 1,3-oxazolyl (oxazolyl), 1,2-oxazolyl(isoxazolyl), oxadiazolyl, 1,3-thiazolyl (thiazolyl), 1,2-thiazolyl(isothiazolyl), tetrazolyl, pyridinyl (pyridyl) pyridazinyl,pyrimidinyl, pyrazinyl, 1,2,3-triazinyl, 1,2,4-triazinyl,1,3,5-triazinyl, 1,2,4,5-tetrazinyl, indazolyl, indolyl,benzothiophenyl, benzofuranyl, benzothiazolyl, benzimidazolyl,benzodioxolyl, acridinyl, quinolinyl, isoquinolinyl, quinazolinyl,quinoxalinyl, phthalazinyl, thienothiophenyl, 1,8-naphthyridinyl, othernaphthyridinyls, pteridinyl or phenothiazine. Where the heteroaryl groupincludes more than one ring, each additional ring is the saturated form(perhydro form) or the partially unsaturated form (for example thedihydro form or tetrahydro form) or the maximally unsaturated(nonaromatic) form. The term heteroaryl thus includes bicyclic radicalsin which the two rings are aromatic and bicyclic radicals in which onlyone ring is aromatic. Such examples of heteroaryl are include3H-indolinyl, 2(1H)-quinolinonyl, 4-oxo-1,4-dihydroquinolinyl,2H-1-oxoisoquinolyl, 1,2-dihydroquinolinyl, (2H)quinolinyl N-oxide,3,4-dihydroquinolinyl, 1,2-dihydroiso-quinolinyl,3,4-dihydro-isoquinolinyl, chromonyl, 3,4-dihydroiso-quinoxalinyl,4-(3H)quinazolinonyl, 4H-chromenyl, 4-chromanonyl, oxindolyl,1,2,3,4-tetrahydro-isoquinolinyl, 1,2,3,4-tetrahydro-quinolinyl,1H-2,3-dihydroisoindolyl, 2,3-dihydrobenzo[f]isoindolyl,1,2,3,4-tetrahydro-benzo[g]isoquinolinyl,1,2,3,4-tetrahydro-benzo[g]isoquinolinyl, chromanyl, isochromanonyl,2,3-dihydro-chromonyl, 1,4-benzo-dioxanyl,1,2,3,4-tetrahydro-quinoxalinyl, 5,6-dihydroquinolyl,5,6-dihydroiso-quinolyl, 5,6-dihydroquinoxalinyl,5,6-dihydroquinazolinyl, 4,5-dihydro-1H-benzimidazolyl,4,5-dihydrobenzoxazolyl, 1,4-naphthoquinolyl,5,6,7,8-tetrahydro-quinolinyl, 5,6,7,8-tetrahydro-isoquinolyl,5,6,7,8-tetrahydroquinoxalinyl, 5,6,7,8-tetrahydroquinazolyl,4,5,6,7-tetrahydro-1H-benzimidazolyl, 4,5,6,7-tetrahydro-benzoxazolyl,1H-4-oxa-1,5-diaza-naphthalen-2-onyl,1,3-dihydro-imidizolo-[4,5]-pyridin-2-onyl,2,3-dihydro-1,4-dinaphthoquinonyl,2,3-dihydro-1H-pyrrol[3,4-b]quinolinyl,1,2,3,4-tetrahydrobenzo[b]-[1,7]naphthyridinyl,1,2,3,4-tetra-hydrobenz-[b][1,6]-naphthyridinyl,1,2,3,4-tetrahydro-9H-pyrido[3,4-b]indolyl,1,2,3,4-tetrahydro-9H-pyrido[4,3-b]indolyl,2,3-dihydro-1H-pyrrolo-[3,4-b]indolyl,1H-2,3,4,5-tetrahydro-azepino[3,4-b]indolyl,1H-2,3,4,5-tetrahydroazepino-[4,3-b]indolyl,1H-2,3,4,5-tetrahydroazepino[4,5-b]indolyl,5,6,7,8-tetrahydro-[1,7]-napthyridinyl,1,2,3,4-tetrahydro-[2,7]-naphthyridyl,2,3-dihydro[1,4]dioxino[2,3-b]pyridyl,2,3-dihydro[1,4]-dioxino[2,3-b]pyridyl,3,4-dihydro-2H-1-oxa[4,6]-diazanaphthalenyl,4,5,6,7-tetrahydro-3H-imidazo[4,5-c]pyridyl,6,7-dihydro[5,8]-diazanaphthalenyl,1,2,3,4-tetrahydro[1,5]-napthyridinyl,1,2,3,4-tetrahydro[1,6]-napthyridinyl,1,2,3,4-tetrahydro[1,7]-napthyridinyl,1,2,3,4-tetrahydro-[1,8]-napthyridinyl or1,2,3,4-tetrahydro[2,6]-napthyridinyl. In some embodiments, heteroarylgroups are optionally substituted. In one embodiment, the one or moresubstituents are each independently selected from among halo, hydroxy,amino, cyano, nitro, alkylamido, acyl, C₁₋₆-alkoxy, C₁₋₆-alkyl,C₁₋₆-haloalkyl, C₁₋₆-hydroxyalkyl, C₁₋₆-aminoalkyl, C₁₋₆-alkylamino,alkylsulfenyl, alkylsulfinyl, alkylsulfonyl, sulfamoyl, ortrifluoromethyl.

Examples of heteroaryl groups include, but are not limited to,unsubstituted and mono- or di-substituted derivatives of furan,benzofuran, thiophene, benzothiophene, pyrrole, pyridine, indole,oxazole, benzoxazole, isoxazole, benzisoxazole, thiazole, benzothiazole,isothiazole, imidazole, benzimidazole, pyrazole, indazole, tetrazole,quinoline, isoquinoline, pyridazine, pyrimidine, purine and pyrazine,furazan, 1,2,3-oxadiazole, 1,2,3-thiadiazole, 1,2,4-thiadiazole,triazole, benzotriazole, pteridine, phenoxazole, oxadiazole,benzopyrazole, quinolizine, cinnoline, phthalazine, quinazoline andquinoxaline. In some embodiments, the substituents are halo, hydroxy,cyano, O—C₁₋₆-alkyl, C₁₋₆-alkyl, hydroxy-C₁₋₆-alkyl andamino-C₁₋₆-alkyl.

As used herein, the term “arylalkyl” alone or in combination, refers toan alkyl substituted with an aryl that can be optionally substituted.

As used herein, the term “heteroarylalkyl” alone or in combination,refers to an alkyl substituted with a heteroaryl that is optionallysubstituted.

As used herein, the term “optionally substituted,” refers to a group inwhich none, one, or more than one of the hydrogen atoms has beenreplaced with one or more group(s) individually and independentlyselected from among alkyl, cycloalkyl, heteroalkyl, aryl, heteroaryl,non-aromatic heterocycle, hydroxy, alkoxy, aryloxy, mercapto, alkylthio,arylthio, cyano, halo, carbonyl, thiocarbonyl, O-carbamyl, N-carbamyl,O-thiocarbamyl, C-amido, N-thiocarbamyl, N-amido, S-sulfonamido,N-sulfonamido, C-carboxy, O-carboxy, isocyanato, thiocyanato,isothiocyanato, nitro, silyl, trihalomethanesulfonyl, and amino,including mono and di substituted amino groups, and the protectedderivatives of amino groups. Such protective derivatives (and protectinggroups that can form such protective derivatives) are known to those ofskill in the art and can be found in references such as Greene and Wuts(Protective Groups in Organic Synthesis, 3^(rd) ed., John Wiley & Sons,New York, N.Y., 1999), which is incorporated herein in its entirety. Inembodiments in which two or more hydrogen atoms have been substituted,the substituent groups can together form a ring.

As used herein, the term “non-aromatic heterocycle” refers to anon-aromatic ring wherein one or more atoms forming the ring is aheteroatom. Non-aromatic heterocyclic rings can be formed by three,four, five, six, seven, eight, nine, or more than nine atoms.Non-aromatic heterocycles can be optionally substituted. In certainembodiments, non-aromatic heterocycles contain one or more carbonyl orthiocarbonyl groups such as, for example, oxo- and thio-containinggroups. Examples of non-aromatic heterocycles include, but are notlimited to, lactams, lactones, cyclic imides, cyclic thioimides, cycliccarbamates, tetrahydrothiopyran, 4H-pyran, tetrahydropyran, piperidine,1,3-dioxin, 1,3-dioxane, 1,4-dioxin, 1,4-dioxane, piperazine,1,3-oxathiane, 1,4-oxathiine, 1,4-oxathiane, tetrahydro-1,4-thiazine,2H-1,2-oxazine, maleimide, succinimide, barbituric acid, thiobarbituricacid, dioxopiperazine, hydantoin, dihydrouracil, morpholine, trioxane,hexahydro-1,3,5-triazine, tetrahydrothiophene, tetrahydrofuran,pyrroline, pyrrolidine, pyrrolidone, pyrrolidione, pyrazoline,pyrazolidine, imidazoline, imidazolidine, 1,3-dioxole, 1,3-dioxolane,1,3-dithiole, 1,3-dithiolane, isoxazoline, isoxazolidine, oxazoline,oxazolidine, oxazolidinone, thiazoline, thiazolidine and1,3-oxathiolane.

Throughout the specification, groups and substituents thereof can bechosen by one skilled in the field to provide stable moieties andcompounds.

It is to be understood that the compounds provided herein can containchiral centers. Such chiral centers can be of either the (R) or (S)configuration, or can be a mixture thereof. Thus, the compounds providedherein can be enantiomerically pure or be stereoisomeric ordiastereomeric mixtures.

As used herein, “enantiomer” refers to one of a pair of molecularentities that are mirror images of each other and non-superimposable.Enantiomeric excess (ee) can be calculated for a mixture of (R) and(S)-enantiomers. The ee can be defined as the absolute value of the molefractions of F_((R)) minus the mole fraction of F_((S)). The percent eethen is the absolute value of the mole fractions of F_((R)) minus themole fraction of F_((S)) multiplied by 100.

As used herein, the term “substantially pure” means sufficientlyhomogeneous to appear free of readily detectable impurities asdetermined by standard methods of analysis, such as thin layerchromatography (TLC), gel electrophoresis, high performance liquidchromatography (HPLC) and mass spectrometry (MS), used by those of skillin the art to assess such purity, or sufficiently pure such that furtherpurification would not detectably alter the physical and chemicalproperties, such as enzymatic and biological activities, of thesubstance. Thus, substantially pure object species (e.g., compound) isthe predominant species present (i.e., on a molar basis it is moreabundant than any other individual species in the composition). Incertain embodiments, a substantially purified fraction is a compositionwherein the object species includes at least about 50 percent (on amolar basis) of all species present. In certain embodiments, asubstantially pure composition will include more than about or 50%, 60%,70%, 80%, 85%, 90%, 95%, or 99% of all species present in thecomposition. In certain embodiments, a substantially pure compositionwill include more than about or 80%, 85%, 90%, 95%, or 99% of allspecies present in the composition. Methods for purification of thecompounds to produce substantially chemically pure compounds are knownto those of skill in the art. A substantially chemically pure compoundcan, however, be a mixture of stereoisomers. In such instances, furtherpurification might increase the specific activity of the compound. Theinstant disclosure is meant to include all such possible isomers, aswell as, their racemic and optically pure forms. Optically active (+)and (−), (R)- and (S)-, or (D)- and (L)-isomers can be prepared usingchiral synthons or chiral reagents, or resolved using conventionaltechniques, such as reverse phase HPLC. When the compounds describedherein contain olefinic double bonds or other centers of geometricasymmetry, and unless specified otherwise, it is intended that thecompounds include both E and Z geometric isomers. Likewise, alltautomeric forms also are intended to be included.

The compounds described herein can be administered alone or incombination with one or more than one other active ingredient or drug oragent. The individual components of the combination can be administeredseparately at different times during the course of therapy orconcurrently in divided or single combination forms. As used herein, theterm “co-administer” refers to administering more than onepharmaceutical agent to a subject. In certain embodiments,co-administered pharmaceutical agents are administered together in asingle dosage unit. In certain embodiments, co-administeredpharmaceutical agents are administered separately. In certainembodiments, co-administered pharmaceutical agents are administered atthe same time. In certain embodiments, co-administered pharmaceuticalagents are administered at different times. The methods disclosed hereintherefore are to be understood as embracing all such regimes ofsimultaneous or alternating treatment.

As used herein, “PO” refers to Per Os, meaning by mouth or orally.

As used herein, the term “subject” is an animal, typically a mammal,including human.

As used herein, the term “patient” includes human and animal subjects.

As used herein, the term “tissue-selective” refers to the ability of acompound to modulate a biological activity in one tissue to a greater orlesser degree than it modulates a biological activity in another tissue.The biological activities in the different tissues can be the same orthey can be different. The biological activities in the differenttissues can be mediated by the same type of target receptor. In certainembodiments, e.g., a tissue-selective compound can modulate an androgenreceptor mediated biological activity in one tissue and fail tomodulate, or modulate to a lesser degree, an androgen receptor mediatedbiological activity in another tissue type.

As used herein, the term “monitoring” refers to observing an effect orabsence of any effect. In certain embodiments, one monitors cells aftercontacting those cells with a compound provided herein. Examples ofeffects that can be monitored include, but are not limited to, changesin cell phenotype, cell proliferation, androgen receptor activity, orthe interaction between an androgen receptor and a natural bindingpartner.

As used herein, the term “cell phenotype” refers to physical orbiological characteristics. Examples of characteristics that constitutephenotype included, but are not limited to, cell size, cellproliferation, cell differentiation, cell survival, apoptosis, or theutilization of a metabolic nutrient (e.g., glucose uptake). Certainchanges or the absence of changes in cell phenotype are readilymonitored using techniques known in the art.

As used herein, the term “contacting” refers to bringing two or morematerials into close enough proximity whereby they can interact. Incertain embodiments, contacting can be accomplished in a vessel such asa test tube, a petri dish, or the like. In certain embodiments,contacting can be performed in the presence of additional materials. Incertain embodiments, contacting can be performed in the presence ofcells. In certain of such embodiments, one or more of the materials thatare being contacted can be inside a cell. Cells can be alive or can bedead. Cells can or can not be intact.

As used herein, “arthritic condition” or “arthritis” refers to a diseasewhose underlying etiology is inflammation of a joint, usuallyaccompanied by pain, such as osteoarthritis and rheumatoid arthritis(Taber's Cyclopedic Medical Dictionary; 14^(th) edition, 1983). Thecompounds disclosed herein are useful, alone or in combination, to treator prevent arthritic conditions. Exemplary arthritic conditions includeBehcet's disease; bursitis and tendinitis; CPPD deposition disease;carpal tunnel syndrome; Ehlers-Danlos syndrome; fibromyalgia; gout;infectious arthritis; inflammatory bowel disease; juvenile arthritis;lupus erythematosus; Lyme disease; Marfan syndrome; myositis;osteoarthritis; osteogenesis imperfecta; osteonecrosis; polyarteritis;polymyalgia rheumatica; psoriatic arthritis; Raynaud's phenomenon;reflex sympathetic dystrophy syndrome; Reiter's syndrome; rheumatoidarthritis; scleroderma; and Sjogren's syndrome. (Bijlsma et al., Am JReprod Immunol 28(34): 231-234 (1992); Cutolo et al., Ann. N.Y. Acad.Sci. 966: 131-142 (2002); Cutolo, Rheum Dis Clin North Am 26(4): 881-895(2000); Jansson et al., Arthritis Rheum 44(9): 2168-2175 (2001); MerckManual (17^(th) edition, pp. 449-451) and Purdie, Br Med Bull 56(3):809-823 (2000)).

As used herein, “NSAIDs” refer to non-steroidal anti-inflammatory drugs.These drugs exhibit anti-inflammatory and analgesic effects and arecommonly used to reduce inflammation and pain, including by decreasingprostaglandin production. Exemplary NSAIDs include, but are not limitedto, aspirin, diclofenac/misoprostol, diclofenac potassium, diclofenac,diflunisal, etodolac, fenoprofen, flurbiprofen, ibuprofen, indomethacin,ketoprofen, meclofenamate, mefanamic acid, meloxicam, nabumetone,naproxen and naproxen sodium, oxaprozin, piroxicam, sodium sulindac andtolmetin.

As used herein, “COX-2 inhibitors” refers to drugs that inhibit only theinducible form of the Cyclooxygenase (COX) enzyme (EC 1.14.99.1), whichis referred to as COX-2. These compounds are well known in the art(e.g., see PNAS, 89: 7384 (1992); Arch. Opthalmol. 108: 573 (1990); FEBSLetters 372: 83 (1995); Clin. Orthop. 313: 76 (1995); J. Mol.Endocrinol. 16: 107 (1996); Cancer Res. 57: 1625 (1997); Cell 93: 705(1998); Intl. J. Mol. Med. 2: 715 (1998) and J. Biol. Chem. 274: 9116(1999)). Exemplary COX-2 inhibitors include, but are not limited to,celecoxib, rofecoxib and valdecoxib.

As used herein, “DMARDs (Disease-Modifying Anti-Rheumatic Drugs)” referto drugs that function by acting upon the immune system of a subject toslow or stop the underlying processes that cause certain forms ofinflammatory arthritis, including rheumatoid arthritis (RA), ankylosingspondylitis, and psoriatic arthritis. DMARDs have been shown to beeffective in the treatment of rheumatoid arthritis, psoriatic arthritis,and ankylosing spondylitis and, for some subjects, these drugs can stopprogression of the disease. Exemplary DMARDs include, but are notlimited to, adalimumab, leflunomide, auranofin, sodium aurothiomalate,chloroquine, etanercept, infliximab, sulfasalazine, mycophenolate,myochrysine, cyclosporine, cyclophosphamide, azathioprine, chlorambucil,methotrexate, minocycline, penicillamine and hydroxychloroquine.

As used herein, “photodynamic therapy dyes” refers to any of theconjugated or unconjugated dyes or pigments used in photodynamictherapy. The photodynamic therapy dyes include porphyrins, chlorines,purpurins, benzoporphyrins and other dyes and pigments that absorb lightof a particular wavelength, thereby initiating tumor necrosis presumablythrough formation of singlet oxygen or other destructive chemicalspecies. Photodynamic therapy for the treatment of cancer is well knownin the art (e.g., see U.S. Pat. Nos. 7,018,395, 7,011,812, 6,806,284,6,723,750, 6,710,066 and 6,630,128).

As used herein, “toxins and cytotoxic drugs” refers to chemicalmolecules that are known to affect the growth and action of some cells,and in some instances leads to the death of the cell when the cells areexposed to the chemical molecules. Exemplary toxins and cytotoxic agentsinclude, but are not limited to, adrenocortical suppressants, such asmitotane; alkyl sulfonates, such as busulfan; ethylenimine derivatives,such as thiotepa; nitrosoureas, such as carmustine, lomustine, semustineand streptozocin; folic acid analogs, such as methotrexate; methylhydrazine derivatives, such as procarbazine; nitrogen mustards, such asmechlorethamine, cyclophosphamide, melphalan, uracil mustard andchlorambucil; purine analogs, such as mercaptopurine and thioguanine;pyrimidine analogs, such as fluorouracil, cytarabine and azaribine;substituted urea compounds, such as hydroxyurea; taxol; triazenes, suchas dacarbazine; and vinca alkaloids, such as vinblastine andvincristine.

As used herein, an “anti-proliferative agent” refers to any agent thatreduces the rate or level of cellular proliferation. The agent canreduce proliferation by inducing apoptosis, modulating cellularmicrotubule structure (e.g., promoting microtubule polymerization),inhibiting tyrosine kinase mediated signaling, antagonizing cell surfacereceptor binding (e.g., EGFR and VEGFR inhibitors), modulatingglucocorticoid receptor functioning, down-regulating angiogenesis (e.g.,inhibiting VEGF functioning) or inducing cell death.

As used herein, an “anti-tumor agent” refers to any agent that limitsthe growth of or destroys a tumor, and includes the following classes ofcompounds: angiogenesis inhibitors, DNA intercalators/cross-linkers, DNAsynthesis inhibitors, DNA-RNA transcription regulators, enzymeinhibitors, gene regulators and microtubule inhibitors.

As used herein, “angiogenesis inhibitors” refers to compounds thatinhibit the formation of new blood vessels, regardless of mechanism.Examples of angiogenesis inhibitors include, but are not limited to,tyrosine kinase inhibitors, such as inhibitors of the tyrosine kinasereceptors Flt-1 (VEGFR1) and Flk-1/KDR (VEGFR2), inhibitors ofepidermal-derived, fibroblast-derived, or platelet derived growthfactors, MMP (matrix metalloprotease) inhibitors, integrin blockers,interferon-α, interleukin-12, pentosan polysulfate, cyclooxygenaseinhibitors, including nonsteroidal anti-inflammatories (NSAIDs) likeaspirin and ibuprofen as well as selective cyclo-oxygenase-2 inhibitorslike celecoxib and rofecoxib, steroidal anti-inflammatories (such ascorticosteroids, mineralocorticoids, dexamethasone, prednisone,prednisolone, methylpred, betamethasone), carboxyamidotriazole,combretastatin A4, squalamine, 6-O-chloroacetyl-carbonyl)-fumagillol,thalidomide, angiostatin, troponin-1, angiotensin II antagonists (seeFernandez et al., J. Lab. Clin. Med. 105: 141-145 (1985)), andantibodies to VEGF (see, Nature Biotechnology 17: 963-968 (October1999); Kim et al., Nature 362: 841-844 (1993), WO 00/44777 and WO00/61186). Other therapeutic agents that modulate or inhibitangiogenesis include agents that modulate or inhibit the coagulation andfibrinolysis systems (see review in Clin. Chem. La. Med. 38: 679-692(2000)). Examples of such agents that modulate or inhibit thecoagulation and fibrinolysis pathways include, but are not limited to,heparin (see Thromb. Haemost. 80: 10-23 (1998)), low molecular weightheparins and carboxypeptidase U inhibitors (also known as inhibitors ofactive thrombin activatable fibrinolysis inhibitor [TAFIa]) (seeThrombosis Res. 101: 329-354 (2001)).

As used herein, “HMG-CoA reductase inhibitors” refers to inhibitors of3-hydroxy-3-methylglutaryl-CoA reductase. Exemplary HMG-CoA reductaseinhibitors include, but are not limited to, lovastatin (see U.S. Pat.Nos. 4,231,938, 4,294,926 and 4,319,039), simvastatin (see U.S. Pat.Nos. 4,444,784, 4,820,850 and 4,916,239), pravastatin (see U.S. Pat.Nos. 4,346,227, 4,537,859, 4,410,629, 5,030,447 and 5,180,589),fluvastatin (see U.S. Pat. Nos. 5,354,772, 4,911,165, 4,929,437,5,189,164, 5,118,853, 5,290,946 and 5,356,896), atorvastatin (see U.S.Pat. Nos. 5,273,995, 4,681,893, 5,489,691 and 5,342,952) andcerivastatin, also known as rivastatin (see U.S. Pat. No. 5,177,080).The structural formulas of these and additional HMG-CoA reductaseinhibitors are described at page 87 of M. Yalpani, “Cholesterol LoweringDrugs,” Chemistry & Industry, pp. 85-89 and U.S. Pat. Nos. 4,782,084 and4,885,314. The term HMG-CoA reductase inhibitor as used herein includesall pharmaceutically acceptable lactone and open-acid forms (i.e., wherethe lactone ring is opened to form the free acid) as well as salt andester forms of compounds that have HMG-CoA reductase inhibitoryactivity.

As used herein, a “synthetic estrogen” refers to non-naturally occurringcompounds that exhibit at least some of the properties of 17β-estradiolwhen administered to a subject. Exemplary synthetic estrogens includeethinyl estradiol, diethylstilbestrol (DES), chlorotrianisene,dienestrol, ethinyl estradiol and ethinyl estradiol 3-cyclopentyl ether.

As used herein, the term “estrogen receptor agonist” refers to acompound that acts at an estrogen receptor and has at least some of thesame biological effects as 17β-estradiol. Compounds that act at anestrogen receptor to block the effects of 17β-estradiol are called“estrogen receptor antagonists.” Compounds exhibiting such selectivityare termed “selective estrogen receptor modulators” or “SERMs.”Exemplary SERMs include bazedoxifene, clomifene, fulvestrant,lasofoxifene, raloxifene, tamoxifen and toremifene.

As used herein, the term “osteoporosis” refers to the conditioncharacterized by reduced bone mass and disruption of bone architecture,resulting in increased bone fragility and increased fracture risk, anddecreased calcification or density of bone. Osteoporosis is a thinningof the bones with reduction in bone mass due to depletion of calcium andbone protein. In osteoporotic patients, bone strength is abnormal, witha resulting increase in the risk of fracture. The fracture can be in theform of cracking (as in a hip fracture) or collapsing (as in acompression fracture of the spine). The spine, hips, and wrists arecommon areas of osteoporosis-induced bone fractures, although fracturesalso can occur in other skeletal areas. Unchecked osteoporosis can leadto changes in posture, physical abnormality and decreased mobility.Osteoporosis can be identified by bone mineral density measurements.

As used herein, “osteopenia” refers to decreased calcification ordensity of bone.

As used herein, a “cathepsin inhibitor” refers to an inhibitor ofcysteine protease. Cysteine proteases, such as cathepsins, are linked toa number of disease conditions, including arthritis, bone remodeling,inflammation and tumor metastasis. Cathepsin protease inhibitors caninhibit osteoclastic bone resorption by inhibiting the degradation ofcollagen fibers and thus are useful in the treatment of bone resorptiondiseases, such as osteoporosis. Examples of cathepsin inhibitors aredescribed in Deaton, Current Topics in Medicinal Chemistry 5(16):1639-1675 (2005), in U.S. Pat. Nos. 7,279,478, 7,279,472, 7,112,589 and7,012,075, and in WO 01/49288 and WO 01/77073.

As used herein, a “proton pump inhibitor” refers to osteoclast vacuolarATPase inhibitors. The proton ATPase found on the apical membrane of theosteoclast has been reported to play a significant role in the boneresorption process and is a target for the design of inhibitors of boneresorption, thereby useful for the treatment and prevention ofosteoporosis and related metabolic diseases (e.g., see Niikura, DrugNews Perspect. 19(3): 139-44 (2006), Visentin et al., J Clin Invest106(2): 309-318 (2000) and Niikura et al., Br J of Pharmacology 142:558-566 (2004)). Exemplary inhibitors include bafilomycin A1, SB242784,FR167356, FR177995, FR202126, FR133605 and NiK-12192[4-(5,6-dichloro-1H-indol-2-yl)-3-ethoxy-N-(2,2,6,6-tetramethyl-piperidin-4-yl)-benzamide](Petrangolini et al., J Pharmacol Exp Ther 318 (3): 939-946 (2006).

As used herein, “PPARγ activators” refers to activators of theperoxisome proliferator-activated receptor gamma (PPARγ), which areknown in the art to inhibit osteoclast-like cell formation and boneresorption (e.g., see Okazaki et al., Endocrinology 140: 5060-5065(1999)). Exemplary PPARγ activators include the glitazones, such asciglitazone, darglitazone, englitazone, troglitazone, pioglitazone,rosiglitazone, the thiazolidinediones (see, e.g., Yki-Järvinen, New EngJ Med 351(11): 1106-1118 (2004), netoglitazone, 15deoxy-Δ_(12,14)-prostaglandin J₂ and analogs, derivatives, andpharmaceutically acceptable salts thereof.

As used herein, “muscle wasting” refers to atrophy or loss of muscletissue, which can result from disease or disuse (lack of exercise). Asused herein, muscle wasting also includes loss of muscle tone andneurogenic atrophy. Muscle wasting is characterized by a weakening,shrinking, and loss of muscle tissue, often caused by degradation of thecontractile myofibrillar proteins actin and myosin (e.g., see Hasselgrenet al., Int'l J of Biochemistry & Cell Biology 37(10): 1932 (225); Lynchet al., Pharmacology & Therapeutics 113, (3): 461-487 (2007)).

As used herein, “chronic muscle wasting” refers to chronic (i.e.,persisting over a long period of time) progressive loss of muscle massand/or to the chronic progressive weakening and degeneration of muscle.

As used herein, “cachexia” refers to weakness and a loss of weightcaused by a disease or as a side effect of illness. Cardiac cachexia,which includes muscle protein wasting of both the cardiac and skeletalmuscle, is a characteristic of congestive heart failure. Cancer cachexiais a syndrome that occurs in patients with solid tumors andhematological malignancies and is manifested by weight loss with massivedepletion of both adipose tissue and lean muscle mass. Cachexia alsooccurs in acquired immunodeficiency syndrome (AIDS). Humanimmunodeficiency virus (HIV)-associated myopathy and/or muscleweakness/wasting is a relatively common clinical manifestation of AIDS.Individuals with HIV-associated myopathy or muscle weakness or wastingtypically experience significant weight loss, generalized or proximalmuscle weakness, tenderness, and muscle atrophy.

As used herein, “sarcopenia” refers to a debilitating disease thatafflicts the elderly and chronically ill patients and is characterizedby loss of muscle mass and function.

As used herein, the term “obesity” refers to the state of being wellabove one's normal weight. Traditionally, a person is considered to beobese if they are more than 20 percent over their ideal weight. Obesityhas been defined by the National Institute of Health (NIH) as a Body toMass Index (BMI) of 30 or above. Overweight due to obesity is asignificant contributor to health problems. It increases the risk ofdeveloping a number of diseases including, for example, type 2 diabetes,high blood pressure (hypertension), stroke, heart attack (myocardialinfarction), heart failure, certain forms of cancer, such as prostatecancer and colon cancer, gallstones and gallbladder disease(cholecystitis), gout and gout-related arthritis, osteoarthritis(degenerative arthritis) of the knees, hips, and the lower back, sleepapnea and Pickwickian syndrome (obesity, red face, underventilation anddrowsiness). As used herein, the term “obesity” includes any one of theabove-listed obesity-related conditions and diseases.

As used herein, the term “cancer” or “neoplastic disease” refers to abroad group of malignant neoplasms, growth or tumor caused by abnormaland uncontrolled cell division, and includes carcinomas and sarcomas(Taber's Cyclopedic Medical Dictionary, 14^(th) edition, 1983). Inadditional to their uncontrolled growth, cancer cells invade and destroyadjacent tissues and often metastasize to new sites within the body.

As used herein, the term “treating cancer” or “treatment of cancer”refers to administration of a treatment to a mammal afflicted with acancerous condition and refers to an effect that alleviates thecancerous condition by killing the cancerous cells, as well as an effectthat results in the inhibition of growth and/or metastasis of thecancer.

As used herein, the term “lipid profile” refers to total cholesterol,low density lipoprotein (LDL), high density lipoprotein (HDL), very lowdensity lipoprotein (VLDL), and triglycerides in a subject. LDL, HDL andVLDL are the three types of lipoproteins found in the blood, and theyusually represent the types of cholesterol found in the blood(cholesterol combined with a protein and triglyceride).

As used herein, the term “anemia” refers to the condition of having lessthan the normal number of red blood cells or less than the normalquantity of hemoglobin in the blood. Because of the decreased number ofred blood cells or reduced quantity of hemoglobin, the oxygen-carryingcapacity of the blood is decreased. A subject with anemia may feel tiredand fatigue easily, appear pale, develop palpitations and become usuallyshort of breath. There are many forms of anemia, including aplasticanemia, Fanconi anemia, hereditary spherocytosis, iron deficiencyanemia, osteopetrosis, pernicious anemia, sickle cell disease,thalassemia, myelodysplastic syndrome, and a variety of bone marrowdiseases.

As used herein, the term “depression” refers to an illness that involvesthe body, mood and thoughts, that affects the way a person eats and/orsleeps and the way one feels about oneself and thinks about things. Thesigns and symptoms of depression include loss of interest in activities,loss of appetite or overeating, loss of emotional expression, an emptymood, feelings of hopelessness, pessimism, guilt or helplessness, socialwithdrawal, fatigue, sleep disturbances, trouble concentrating,remembering, or making decisions, restlessness, irritability, headaches,digestive disorders or chronic pain.

As used herein, the term “sexual dysfunction” refers to impairment ofthe emotional or physical responses associated with sexual activity,including sexual desire disorders, sexual arousal disorders, orgasmdisorders, and sexual pain disorders, which can prevent an individualfrom engaging in sexual activity or result in inadequate sexualfunctioning. Sexual dysfunction includes lack of sexual desire, anxietyabout sexual performance, difficulty in becoming aroused, inability toachieve orgasm (anorgasmia), premature ejaculation, erectiledysfunction, impotence, frigidity, dyspareunia, vaginismus anddyspareunia (e.g., see American Society for Reproductive Medicine,“Sexual Dysfunction—Patient's Fact Sheet” (1998)).

As used herein, the term “male sexual dysfunction” includes impotence,loss of libido, orgasm dysfunction (e.g., premature ejaculation orretrograde ejaculation) and erectile dysfunction.

As used herein, the term “erectile dysfunction” refers to a disorderinvolving the failure of a male mammal to achieve erection, ejaculation,or both. Symptoms of erectile dysfunction include an inability toachieve or maintain an erection, ejaculatory failure, prematureejaculation, or inability to achieve an orgasm. An increase in erectiledysfunction and sexual dysfunction can have numerous underlying causes,including but not limited to (1) aging, (b) an underlying physicaldysfunction, such as trauma, surgery, and peripheral vascular disease,and (3) side-effects resulting from drug treatment, depression, andother CNS disorders.

As used herein, the term “female sexual dysfunction” includesdysfunction in desire, sexual arousal, sexual receptivity, and orgasmrelated to disturbances in the clitoris, vagina, periurethral glans, andother trigger points of sexual function. In particular, anatomic andfunctional modification of such trigger points may diminish the orgasmicpotential in breast cancer and gynecologic cancer patients. Treatment offemale sexual dysfunction with an SARM compound provided herein canresult in improved blood flow, improved lubrication, improved sensation,facilitation of reaching orgasm, reduction in the refractory periodbetween orgasms, and improvements in arousal and desire.

As used herein, the term “libido” refers to sexual desire.

As used herein, the term “hypogonadism” refers to a condition resultingfrom or characterized by abnormally decreased functional activity of thegonads, with retardation of growth and sexual development.

As used herein, the term “cognition” refers to the process of knowing,specifically the process of being aware, knowing, thinking, learning andjudging. As used herein, the term “mood” refers to a temper or state ofthe mind. As used herein, the term “alteration” or “alterations” refersto any change for the positive or negative, in cognition and/or mood.

As used herein, the term “hair loss” refers to alopecia, or baldness,such as in the common type of male-pattern baldness. Hair loss affectsboth males and females.

As used herein, “frailty” refers to an adverse, primarily gerontologic,health condition, characterized by low functional reserve, acceleratedosteoporosis, easy tiring, decreased muscle strength, highsusceptibility to disease and decreased libido (e.g., see Bandeen-Rocheet al., The Journals of Gerontology Series A: Biological Sciences andMedical Sciences 61: 262-266 (2006)).

As used herein, “bioavailability” refers to the rate and extent to whichthe active substance or therapeutic moiety is absorbed from apharmaceutical form and becomes available at the site of action orreaches systemic circulation. The “absolute bioavailability” of a givenpharmaceutical form is compared to that following intravenousadministration, which is by definition 100%. Administration by a routeother than intravenous administration generally is less than 100%, dueto slow or incomplete absorption, or metabolic destruction. “Goodbioavailability” generally is >50% and “poor bioavailability” generallyis <20%.

As used herein, “androgenic activity” refers to androgen receptor (AR)agonist activity in androgenic target tissues, such as prostate andseminal vesicles. Androgenic activity is typically demonstrated byincreases in the weights of the prostate and seminal vesicles, which areaccepted in the art as indicators of androgenic activity (e.g., seeLemus et al., J Steroid Biochem Mol Biol 60(1-2): 121-129 (1997)).

As used herein, “androgenic effect” or “androgenic effects” refers toproducing or enhancing male traits, and includes producing side effectsassociated with administration of steroidal androgens such astestosterone. These adverse androgenic effects include manifestationssuch as prostate enlargement, acne, repression of high densitylipoprotein cholesterol (HDL), hirsutism, virilization andmasculinization.

As used herein, “anabolic activity” refers to increasing the mass and/orstrength of a tissue, such as a connective tissue. Increases in theweight of the levator ani muscle are indicative of anabolic activity,and are accepted in the art as a reliable index of anabolic activity(e.g., see Antonio et al., J Appl Physiol 87: 2016-2019 (1999)).Anabolic activity in bone and muscle decreases bone fracture rates in asubject. Anabolic activity of the compounds provided herein on musclecan be tested by assessing expression of MHC subtypes in skeletal muscle(e.g., see Wright et al., J Appl Physiol. 83(4): 1389-96 (1997)). Boneformation rate, another indication of anabolic activity, can be assessedby osteocalcin level measurement. Plasma osteocalcin levels can bedetermined using any method known in the art (e.g., see Koyama et al., JImmunol Methods 139(1): 17-23 (1991)). A rat osteocalcin EIA kit iscommercially available from Biomedical Technologies Inc. (Stoughton,Mass.).

As used herein, “connective tissue” refers to tissue generally ofmesodermal origin that is characterized by a highly vascular matrix andwhich forms the supporting and connecting structures of the body.Connective tissue includes collagenous, elastic, and reticular fibers,muscle, adipose tissue, cartilage, and bone. Exemplary connective tissueincludes adipose tissue, areolar tissue, blood, bone (includingcancellous bone, compact bone, cortical bone, spongy bone and trabecularbone), bone marrow, cartilage, collagen, cutis, elastic tissue,endoneurium, fascia, ligament, mesenchymal connective tissue, mucousconnective tissue, muscle, osseous tissue, perineurium, perimysium,submucosa and tendon.

As used herein, “bone mineral density” or “BMD” refers to the density ofminerals (such as calcium) in bone. BMD is determined using a specialX-ray, computed tomography (CT) scan, or ultrasound. This information isused to estimate the strength of bones. Increasing mineral content ofbone increases the density of the bone and its strength. The denser thebone, the less likely it is to break.

As used herein, “unit dosage forms” refers to physically discrete unitssuitable for human and animal subjects. Each unit dosage includes apredetermined quantity of the therapeutically active compound sufficientto produce the desired therapeutic effect, in association with, whenrequired, a pharmaceutical carrier, vehicle or diluent. Examples of unitdosage forms include tablets, capsules, pills, powders, granules,sterile parenteral solutions or suspensions, ampoules and syringes, andoral solutions or suspensions, and oil-water emulsions. Unit dosageforms can be individually packaged as is known in the art, such as inblister packs. Unit dosage forms can be administered in fractions ormultiples thereof.

As used herein, “multiple dosage forms” refers to a plurality ofidentical unit dosage forms packaged in a single container to beadministered in segregated unit dosage form. Examples of multiple dosageforms include vials, bottles of tablets or capsules or bottles of pintsor gallons of liquid containing the active compound. Hence, a multipledosage form is a multiple of unit dosages which are not segregated inpackaging.

As used herein, “non-specific binding” refers to that binding remainingin the presence of an excess of unlabeled specific ligand (i.e., in thecase of androgen receptor, 1000 nM of unlabeled dihydrotestosterone).

As used herein, “potency” refers to the dose of drug required to producea specific effect of given intensity as compared to a standardreference.

As used herein, the term “AUC” refers to the area under the plasmaconcentration-time curve and can be used as a metric for extent ofexposure of a pharmaceutical.

As used herein, the term “exposure” refers to AUC and the term “totalexposure” refers to AUC_(0-∞).

As used herein, the term “AUC_(0-∞)” or “AUC_(0-inf)” refers to the areaunder the concentration-time curve from time zero to infinity(extrapolated). AUC_(0-∞) can be calculated as AUC_(0-t)+(C_(t)/K_(el))where C_(t) is the calculation at time t.

As used herein, the term “AUC_(0-t)” refers to the area under theconcentration-time curve from time zero to time of last non-zero (lastmeasurable) concentration. The linear trapezoidal rule can be used tocalculate AUC_(0-t).

As used herein, the term “AUC_(x)” refers to the area under theconcentration-time curve from time zero to x hours post dose. Thus, AUC₆refers to the area under the concentration-time curve from time zero to6 hours post-administration and AUC₂₄ refers to the area under theconcentration-time curve from time zero to 24 hours post-administration.

As used herein, the term “C_(MAX)” refers to the maximum (peak) observedplasma concentration.

As used herein, the term “C_(MIN)” refers to the minimum observed plasmaconcentration, which also can be referred to as the troughconcentration.

As used herein, the term “T_(MAX)” refers to the time to reach themaximum (peak) observed plasma concentration C_(MAX).

As used herein, the term “apparent half-life” or “t_(1/2)” refers to theapparent time required for half the quantity of a drug or othersubstance administered to an organism to be metabolized or eliminated.

B. COMPOUNDS

Provided herein are compounds that have a structure of Formula I orFormula II or Formula III:

where R¹ is halogen, pseudohalogen, optionally substituted lower alkyl,optionally substituted haloalkyl or NO₂; R² is hydrogen, halogen,pseudohalogen, optionally substituted lower alkyl or optionallysubstituted lower haloalkyl; R³ is hydrogen, halogen, pseudohalogen,optionally substituted lower alkyl or optionally substituted lowerhaloalkyl; R⁴ is halogen or lower haloalkyl; and R⁵ is lower alkyl orlower haloalkyl; and pharmaceutically acceptable salts, esters andprodrugs thereof.

In one embodiment, R¹ is lower haloalkyl or halogen; R² is hydrogen orlower alkyl; R³ is hydrogen or lower alkyl; R⁴ is CF₃ or halogen; and R⁵is C₁ to C₄ alkyl or C₁ to C₄ haloalkyl; and pharmaceutically acceptablesalts, esters and prodrugs thereof.

In one embodiment, R¹ is CF₃, F, or Cl; R² is H or methyl; R³ is H ormethyl; R⁴ is Cl or CF₃; and R⁵ is methyl, ethyl or CF₃; andpharmaceutically acceptable salts, esters and prodrugs thereof.

For any and all of the embodiments, substituents can be selected fromamong a subset of the listed alternatives.

In certain embodiments, R¹ is CF₃. In certain embodiments, R¹ is F orCl. In certain embodiments, R¹ is F. In certain embodiments, R¹ is Cl.

In certain embodiments, R² is H. In certain embodiments, R² is methyl.

In certain embodiments, R³ is H. In certain embodiments, R³ is methyl.

In certain embodiments, R⁴ is Cl. In certain embodiments, R⁴ is CF₃.

In certain embodiments, R⁵ is methyl or ethyl. In certain embodiments,R⁵ is methyl. In certain embodiments, R⁵ is ethyl. In certainembodiments, R⁵ is CF₃.

The compounds provided herein include:

-   R,R-4-(2-(1-Hydroxyl-2,2,2-trifluoroethyl)pyrrolidinyl)-2-trifluoromethylbenzonitrile;-   4-(2(R)-(1(S)-hydroxyl-2,2,2-trifluoroethyl)pyrrolidinyl)-2-trifluoromethyl-benzonitrile;-   R,R,R-4-(2-(1-Hydroxyl-2,2,2-trifluoroethyl)-5-methylpyrrolidinyl)-2-trifluoromethyl-benzonitrile;-   R,R,R-4-(2-(1-Hydroxyl-2,2,2-trifluoroethyl)-5-methylpyrrolidinyl)-2-chlorobenzonitrile;-   4-(2(R)-(1(S)-hydroxyl-2,2,2-trifluoroethyl)-5(R)-methylpyrrolidinyl)-2-trifluoro-methylbenzonitrile;-   4-(2(R)-(1(S)-hydroxyl-2,2,2-trifluoroethyl)-5(R)-methylpyrrolidinyl)-2-chlorobenzonitrile;-   R,R-4-(2-(1-Hydroxyl-2,2,2-trifluoroethyl)pyrrolidinyl)-2-chlorobenzonitrile;-   R,R-4-(2-(1-Hydroxyl-2,2,2-trifluoroethyl)pyrrolidinyl)-2-chloro-3-methyl-benzonitrile;-   4-(2(R)-(1(S)-hydroxyl-2,2,2-trifluoroethyl)pyrrolidinyl)-2-chlorobenzonitrile;-   4-(2(R)-(1(S)-hydroxyl-2,2,2-trifluoroethyl)pyrrolidinyl)-2-chloro-3-methyl-benzonitrile;-   3-methyl-4-((R)-2-((R)-2,2,2-trifluoro-1-hydroxyethyl)pyrrolidin-1-yl)-2-(trifluoro-methyl)benzonitrile;-   3-methyl-4-((R)-2-((S)-2,2,2-trifluoro-1-hydroxyethyl)pyrrolidin-1-yl)-2-(trifluoro-methyl)benzonitrile;-   3-methyl-4-((2R,5R)-2-methyl-5-((S)-2,2,2-trifluoro-1-hydroxyethyl)pyrrolidin-1-yl)-2-(trifluoromethyl)benzonitrile;-   2-fluoro-4-((2R,5R)-2-methyl-5-((S)-2,2,2-trifluoro-1-hydroxyethyl)pyrrolidin-1-yl)benzonitrile;-   2-fluoro-3-methyl-4-((2R,5R)-2-methyl-5-((S)-2,2,2-trifluoro-1-hydroxyethyl)-pyrrolidin-1-yl)benzonitrile;-   2-fluoro-3-methyl-4-((R)-2-((S)-2,2,2-trifluoro-1-hydroxyethyl)pyrrolidin-1-yl)-benzonitrile;-   2-fluoro-4-((R)-2-(S)-2,2,2-trifluoro-1-hydroxyethyl)pyrrolidin-1-yl)benzonitrile;-   2-chloro-4-((2R,5R)-2-methyl-5-((S)-2,2,2-trifluoro-1-hydroxyethyl)pyrrolidin-1-yl)benzonitrile;-   2-chloro-3-methyl-4-((2R,5R)-2-methyl-5-((S)-2,2,2-trifluoro-1-hydroxyethyl)-pyrrolidin-1-yl)benzonitrile;-   2-chloro-3-methyl-4-((R)-2-((S)-2,2,2-trifluoro-1-hydroxyethyl)pyrrolidin-1-yl)-benzonitrile;-   2-chloro-4-((R)-2-((S)-2,2,2-trifluoro-1-hydroxyethyl)pyrrolidin-1-yl)benzonitrile;-   (3R)-10-chloro-3-methyl-4-(3,3,3-trifluoro-2(R)-hydroxypropyl)-3,4-dihydro-2H-[1,4]oxazino[2,3-f]quinolin-8(7H)-one;-   (3R)-10-chloro-3-ethyl-4-(3,3,3-trifluoro-2(R)-hydroxypropyl)-3,4-dihydro-2H-[1,4]oxazino[2,3-f]quinolin-8(7H)-one;-   (3R)-10-chloro-4-(3,3,3-trifluoro-2(R)-hydroxypropyl)-3-(trifluoromethyl)-3,4-dihydro-2H-[1,4]oxazino[2,3-f]quinolin-8(7H)-one;-   (3R)-10-chloro-3-methyl-4-(3,3,3-trifluoro-2(S)-hydroxypropyl)-3,4-dihydro-2H-[1,4]oxazino[2,3-f]quinolin-8(7H)-one;-   (3R)-10-chloro-3-ethyl-4-(3,3,3-trifluoro-2(S)-hydroxypropyl)-3,4-dihydro-2H-[1,4]oxazino[2,3-f]quinolin-8(7H)-one;-   (3R)-10-chloro-4-(3,3,3-trifluoro-2(S)-hydroxypropyl)-3-(trifluoromethyl)-3,4-dihydro-2H-[1,4]oxazino[2,3-f]quinolin-8(7H)-one;-   (3R)-3-ethyl-4-(3,3,3-trifluoro-2(R)-hydroxypropyl)-10-(trifluoromethyl)-3,4-dihydro-2H-[1,4]oxazino[2,3-f]quinolin-8(7H)-one;-   (3R)-3-methyl-4-(3,3,3-trifluoro-2(R)-hydroxypropyl)-10-(trifluoromethyl)-3,4-dihydro-2H-[1,4]oxazino[2,3-f]quinolin-8(7H)-one;-   (3R)-4-(3,3,3-trifluoro-2(R)-hydroxypropyl)-3,10-bis(trifluoromethyl)-3,4-dihydro-2H-[1,4]oxazino[2,3-f]quinolin-8(7H)-one;-   (3R)-3-ethyl-4-(3,3,3-trifluoro-2(S)-hydroxypropyl)-10-(trifluoromethyl)-3,4-dihydro-2H-[1,4]oxazino[2,3-f]quinolin-8(7H)-one;-   (3R)-3-methyl-4-(3,3,3-trifluoro-2(S)-hydroxypropyl)-10-(trifluoromethyl)-3,4-dihydro-2H-[1,4]oxazino[2,3-f]quinolin-8(7H)-one;-   (3R)-4-(3,3,3-trifluoro-2(S)-hydroxypropyl)-3,10-bis(trifluoromethyl)-3,4-dihydro-2H-[1,4]oxazino[2,3-f]quinolin-8(7H)-one;-   (R)-10-chloro-3-methyl-4-(2,2,2-trifluoroethyl)-3,4-dihydro-2H-[1,4]oxazino-[2,3-f]-quinolin-8(7H)-one;-   (R)-10-chloro-3-ethyl-4-(2,2,2-trifluoroethyl)-3,4-dihydro-2H-[1,4]oxazino[2,3-f]-quinolin-8(7H)-one;    and-   (R)-10-chloro-4-(2,2,2-trifluoroethyl)-3-(trifluoromethyl)-3,4-dihydro-2H-[1,4]-oxazino[2,3-f]quinolin-8(7H)-one;

and pharmaceutically acceptable salts, esters and/or prodrugs thereof.

These compounds are SARMs in that they exhibit tissue selectivity. Insome embodiments, the target tissue is connective tissue. In someembodiments, the target tissue is muscle. In some embodiments, thetarget tissue is bone. In some embodiments, the compounds providedherein have anabolic activity and promote tissue formation and/orgrowth. In some embodiments, the compounds provided herein candemonstrate full agonist activity in connective tissue without adverseandrogenic effects. In some embodiments, the compounds provided hereincan demonstrate full agonist activity in muscle, resulting in increasedor improved muscle mass and muscle strength without adverse androgeniceffects. In some embodiments, the compounds provided herein candemonstrate full agonist activity in bone, resulting in increased orimproved bone density and strength without adverse androgenic effects.Thus, in some embodiments, the SARM compounds provided herein haveutility in treating conditions that are remediated by anabolic activity(e.g., reversing connective tissue loss).

The compounds provided herein also are androgen receptor selectivebinding compounds, in that they bind to any portion of an androgenreceptor with a greater affinity than they bind to a non-androgenreceptor, such as, but not limited to, a progesterone receptor (PR),estrogen receptor (ER), glucocorticoid receptor (GR), mineralocorticoidreceptor (MR), retinoic acid receptor (RAR), rexinoid receptor (RXR), orperoxisome proliferator-activated receptor (PPAR). The high selectivityfor the androgen receptor means that the compounds are unlikely to havenon-target receptor activity.

For example, compound4-(2(R)-(1(S)-hydroxyl-2,2,2-trifluoroethyl)-pyrrolidinyl)-2-trifluoromethylbenzonitrile(Compound 102) is a potent AR agonist with tissue selectivity and ishighly selective for the AR receptor. It has been shown to be orallybioavailable in multiple animal species, including monkeys, indicatinggood bioavailability in humans. Based on its plasma half-life it can beformulated for administration as a once or twice a day drug. It hasshown good tolerance and safety in a multiple dose toxicology study. Itis negative for genotoxicity in in vitro assays.

Certain compounds provided herein can exist as stereoisomers, includingoptical isomers. The present disclosure is intended to include allstereoisomers and the racemic to mixtures of such stereoisomers as wellas the individual enantiomers that can be separated according to any ofa number of conventional methods that are known in the art. Thesemethods include chiral chromatography, derivatization with a chiralauxiliary followed by separation by chromatography or crystallization,and fractional crystallization of diastereomeric salts.

C. PREPARATION OF THE COMPOUNDS

Provided herein are methods of making androgen receptor modulators offormulae I, II and III. In certain embodiments, the compounds providedherein can be synthesized using the following synthesis schemes. In eachof the Schemes, the R groups correspond to the definitions describedabove.

1. Scheme I—Preparation of Compounds of Formula I

Scheme I describes the preparation of compounds of Formula I (shown asStructure 3 in Scheme I).

In Scheme I, compounds of Structure 1 are either commercially availableor easily prepared by known methods. For example,4-fluoro-2-(trifluoromethyl)-benzonitrile (Alfa Aesar, Ward Hill, Mass.,Cat. No. B20617), 2,4-difluorobenzonitrile (Alfa Aesar, Ward Hill,Mass., Cat. No. A14113) 2-chloro-4-fluorobenzonitrile (Alfa Aesar, WardHill, Mass., Cat. No. A15478) and 2,4-difluoro-3-methylbenzonitrile(Fluorochem Ltd—Wesley Street, Old Glossop, Derbyshire SK13 7RY, Cat.No. 033815) are commercially available.

2-Chloro-4-fluoro-3-methylbenzonitrile can be prepared using methodsknown in the art. For example, 2-Chloro-4-fluorobenzonitrile (1 g) andTMEDA (1.13 mL) in THF (10 mL) were cooled to −78° C., under nitrogen.Sec-butyllithium (1.3M in cyclo-hexane, 8.54 mL) was added over 20 min,keeping the temperature below −70° C. The mixture was then stirred at−78° C. for 2.5 h. Methyl iodide (0.5 mL) was added and the mixtureallowed to warm to 15° C., over 35 min. The reaction was quenched with asaturated aqueous solution of ammonium chloride and the product wasextracted with ethyl acetate. The ethyl acetate was washed with brine,dried (MgSO₄), filtered and concentrated under reduced pressure to givecrude product (1.28 g).

2-Chloro-4-fluoro-3-methylbenzonitrile also can be synthesized by adding3-bromo-2-chloro-6-fluorotoluene (173 mg, 0.78 mmol), zinc cyanide (91mg, 0.78 mmol), tetrakis-(triphenylphosphine)palladium(0) (27 mg, 23Tmol) and DMF (1 mL) to a vial, and after sealing the vial, irradiatingthe mixture for 150 sec at 200° C. in a microwave oven. Diethyl ether(30 ml) is then added and the reaction mixture washed with magnesiumsulphate (4% solution, 3×20 mL) followed by brine (20 mL). The organiclayer is dried and evaporated. The product is further purified by columnchromatography on silica gel using n-heptane/ethyl acetate (9:1) givinga white solid.

4-Fluoro-3-methyl-2-(trifluoromethyl)benzonitrile (Compound 1 of SchemeI with fluoro at position 4) is synthesized by methods known in the art.For example, 4-fluoro-2-(trifluoromethyl)benzonitrile (1.22 g, availablefrom Alfa Aesar, Ward Hill, Mass., Cat. No. B20617) and TMEDA (1.13 mL)in THF (10 mL) are cooled to −78° C., under nitrogen. Sec-butyllithium(1.3M in cyclohexane, 8.54 mL) is added over 20 min., keeping thetemperature below −70° C. The mixture is stirred at −78° C. for 2.5 h.Methyl iodide (0.5 mL) is added and the mixture allowed to warm to 15°C., over 35 min. The reaction is quenched with a saturated aqueoussolution of ammonium chloride and the product is extracted with ethylacetate. The ethyl acetate is washed with brine, dried (MgSO₄), filteredand concentrated under reduced pressure to give the crude product, whichis further purified by column chromatography on silica gel usingn-heptane/ethyl acetate (9:1) giving a white solid.

Compounds of Structures 2 and 4 are prepared from D-proline orD-pyroglutamic acid. For example,(R)-2,2,2-trifluoro-1-(pyrrolidin-2-yl)ethanol (Compound 2, Scheme I,R³=hydrogen) is prepared by oxidation of D-prolinol (Compound 4, SchemeI, R³=hydrogen) to afford D-prolinaldehyde (see e.g., Smith et al.,March's Advanced Organic Chemistry, 6^(th) ed., Wiley, N.J., 2007, p.1715-1728). The resulting D-prolinaldehyde is reacted with CF₃-TMS(trimethyl(trifluoromethyl)-silane) to afford the(R)-2,2,2-trifluoro-1-(pyrrolidin-2-yl)ethanol (see e.g., Prakash etal., J. Am. Chem. Soc. 111: 393, 1989).

2,2,2-trifluoro-1-((2R,5R)-5-methylpyrrolidin-2-yl)ethanol (Compound 2,Scheme I, R³=methyl) is prepared by oxidation of((2R,5R)-5-methylpyrrolidin-2-yl)methanol (Compound 4, Scheme I,R³=methyl) to afford the resulting aldehyde,(2R,5R)-5-methylpyrrolidine-2-carbaldehyde (see e.g., Smith et al.,March's Advanced Organic Chemistry, 6^(th) ed., Wiley, N.J., 2007, p.1715-1728). The resulting aldehyde is reacted with CF₃-TMS(trimethyl(trifluoromethyl)silane) to afford2,2,2-trifluoro-1-((2R,5R)-5-methylpyrrolidin-2-yl)ethanol (see e.g.,Prakash et al., J. Am. Chem. Soc. 111: 393, 1989).

(R)-pyrrolidin-2-ylmethanol (Compound 4, Scheme I, R³=hydrogen) also isknown as D-prolinol, which is available from Sigma-Aldrich, Milwaukee,Wis., Cat. No. 81744. (R)-pyrrolidin-2-ylmethanol is prepared byreducing D-proline with LiAlH₄ (see, Smith et al., March's AdvancedOrganic Chemistry, 6^(th) ed., Wiley, N.J., 2007, p. 1805-1806; seealso, Dei et al., Bioorg. Med. Chem. 11: 3153-3164 (2003)).

((2R,5R)-5-methylpyrrolidin-2-yl)methanol (Compound 4, Scheme I,R³=methyl) is prepared by protecting(R)-(−)-5-(hydroxymethyl)-2-pyrrolidinone (available from Sigma Aldrich,Milwaukee, Wis., Cat. No. 366358) at the nitrogen atom with a suitableprotecting group, such as a Boc protecting group, to afford, forexample, (2R,5R)-tert-butyl-2-formyl-5-methylpyrrolidine-1-carboxylate(see e.g., Wuts et al., Green's Protective Groups in Organic Synthesis,4^(th) ed., Wiley, N.J., 2007, p. 725-727). The resulting compound istreated with MeLi or MeMgX, where X is Cl or Br (see e.g., Smith et al.,March's Advanced Organic Chemistry, 6^(th) ed., Wiley, N.J., 2007, p.1300-1309), followed by hydrogenation (see e.g., Smith et al., March'sAdvanced Organic Chemistry, 6^(th) ed., Wiley, N.J., 2007, p. 1053-1062)and cleavage of the nitrogen protecting group (see e.g., Wuts et al.,Green's Protective Groups in Organic Synthesis, 4^(th) ed., Wiley, N.J.,2007, p. 727-735) to afford ((2R,5R)-5-methylpyrrolidin-2-yl)-methanol.

Palladium catalyzed coupling reaction or base-mediated displacementreaction of compounds of Structure 1 and pyrrolidine derivatives ofStructure 2 provide products of Structure 3 in a mixture of twodiastereomers that are separated to give final products. Alternatively,intermediates of Structure 5 are obtained in the similar reactionconditions using Structure 4 and are oxidized to the aldehydes ofStructure 6. Treatment of the aldehydes with TMS-CF₃ followed byseparation of the diastereomers leads to final products of Formula I(shown as Structure 3 in Scheme I).

2. Preparation of Compounds of Formula II

a. Scheme II—Compounds of Formula II where R⁴ is CF₃

Scheme II describes the preparation of compounds of Formula II where R⁴is CF₃ (shown as Structure 15 in Scheme II). In Scheme II, intermediatesof Structure 14 (R⁵=Me, Et) are prepared from compound 12 by a knownmethod described in U.S. Pat. No. 7,214,690, which is incorporatedherein in its entirety.

The process depicted in Scheme II begins with the Knorr cyclization of aphenylenediamine derivative, for example, 5-chloro-1,3-phenylenediamine(Structure 7), with a β-ketoester, or its corresponding hydrate orhemiacetal, for example ethyl 4,4,4-trifluoroacetoacetate, to afford thecorresponding (1H)-quinolin-2-one (see Jones, Comprehensive HeterocyclicChemistry (Katritzky & Rees, eds., Pergamon, New York, Vol. 2, chap.2.08, pp 421-426 (1984), the disclosure of which is herein incorporatedby reference). Reduction of the halide group can be achieved by chemicalreduction, with, for example, a metal catalyst, for example, 10% Pd—C,in a hydrogen atmosphere, to afford a compound of Structure 8.Conversion of the aniline ring to a phenol ring can be effected bytreatment of Structure 8 with a diazotizing agent, for example, sodiumnitrite in sulfuric acid, to afford a compound of Structure 9.Bromination of the phenol ring with a brominating reagent, for example,N-bromosuccinimide, in the presence of a base, for example,diisopropylamine, affords a compound of Structure 10 (see, e.g.,Fujisaki et al., Bull. Chem. Soc. Jpn. 66: 1576-1579 (1993), thedisclosure of which is herein incorporated by reference.

Selective protection of the phenolic oxygen can be achieved by treatmentof Structure 10 with an alkyl halide, for example, benzyl bromide, inthe presence of a base, for example, cesium fluoride, to afford thecorresponding ether. Protection of the pyridone ring, with, for exampleisopropyl iodide, mediated by a base, for example, cesium fluoride,affords the corresponding imino ether (Structure 11). Selectivehydrolysis of the phenolic ether can be accomplished by acidichydrolysis, with, for example, a 1:1 mixture of methanesulfonic acid andacetic acid, to afford a compound of Structure 12.

The compound of Structure 14 is prepared by alkylation of the phenolicoxygen of Structure 12 by treatment with a protected amino alcohol 13.For example, (R)—N-t-boc-2-aminoalkan-1-ol (13, R⁵=Me or Et), is reactedwith the compound of Structure 12 under Mitsunobu conditions, forexample, triphenylphosphine and diisopropyl azodicarboxylate, in thepresence of a base, for example, N-methylmorpholine, to afford thecorresponding Mitsunobu product. Removal of the t-butoxycarbonylprotecting group can be accomplished by acidic hydrolysis, with, forexample, trifluoroacetic acid. Closure of the amine to the aromatichalide can be achieved by treatment with a transition metal, for examplePd₂(dba)₃ in the presence of a ligand, for example, BINAP, and a base,for example, sodium t-butoxide, to afford a compound of Structure 14(see, Wagaw et al., J. Am. Chem. Soc. 119: 8451-8458 (1997)).

Alternatively, intermediate 14 is prepared by O-alkylation of compound12 with a chiral trifluoromethyl epoxide followed by acetate formationto give intermediate 16. Palladium catalyzed amination of the bromoaryl16 followed by an intramolecular cyclization yield intermediate 14.N-Alkylation of intermediates of Structure 14 with a chiraltrifluoromethyl epoxide followed by acid hydrolysis gives final productsof Formula II (Structure 15 in Scheme II) in a stereoselective fashion.

b. Scheme III—Compounds of Formula II where R⁴ is Cl

Scheme III describes the preparation of compounds of Formula II where R⁴is Cl (shown as Structure 20 in Scheme III). In Scheme III, treatment of5-chloro-1,3-diaminobenzene (Alfa Aesar, Ward Hill, Mass., Cat. No.L06485) with ethyl malonate followed by hydrogenation to remove the5-chlorine and chloronation with POCl₃ provide intermediate 17.Diazotization followed by hydrolysis, bromination, and isopropyl etherformation give intermediate 18. O-Alkylation of compound 18 followed byan intramolecular amino coupling reaction yields intermediates ofStructure 19. N-alkylation with a chiral epoxide followed by an acidhydrolysis gives final products of Formula II where R⁴ is Cl (Structure20 in Scheme III).

3. Scheme IV—Preparation of Compounds of Formula III

Scheme IV describes the preparation of compounds of Formula III (shownas Structure 20 is Scheme IV).

In Scheme VI, treatment of 5-chloro-1,3-diaminobenzene (Alfa Aesar, WardHill, Mass., Cat. No. L06485) with ethyl malonate followed byhydrogenation to remove the 5-chlorine and chloronation with POCl₃provide intermediate 17. Diazotization followed by hydrolysis,bromination, and isopropyl ether formation give intermediate 18.O-Alkylation of compound 18 followed by an intramolecular amino couplingreaction yields intermediates of Structure 19. Treatment ofintermediates 19 with TFA and sodium borohydride generates compounds ofFormula III (shown as Structure 21 in Scheme IV).

D. CERTAIN INDICATIONS

Androgen therapy has been used to treat a variety of male disorders suchas reproductive disorders and primary or secondary male hypogonadism. Anumber of natural or synthetic AR agonists have been investigated forthe treatment of musculoskeletal disorders, such as bone disease,hematopoietic disorders, neuromuscular disease, rheumatological disease,wasting disease, and for hormone replacement therapy (HRT), such asfemale androgen deficiency. In addition, AR antagonists, such asflutamide and bicalutamide, are used to treat prostate cancer.

Progress of androgen therapy has been limited by the inability toseparate desirable androgenic activities from undesirable ordose-limiting side effects. Recent advances in the development ofselective androgen receptor modulators (SARMs) that exhibit tissueselectivity in targeting the androgen receptor while eliminatingundesired side effects (e.g., see Negro-Vilar, A. JCE&M 54(10): 3459-62(1999); Reid et al., Investigational New Drugs 17: 271-284 (1999)). Forexample, the compound LG120907, a non-steroidal AR antagonist, has beenshown, in rats, to have reduced antagonist effects on the hypothalamicaxis and on libido (reproductive rate) as compared to other clinicallyused AR antagonists, such as Casodex, and has been characterized as aselective androgen receptor modulator for the treatment of prostatecancer (e.g., see Gao et al., Pharmaceutical Research 23(8): 1641-1658(2006)). Other SARMs and uses thereof have been identified in the art(e.g., see U.S. Pat. Appl. No. US2007254875 and U.S. Pat. Nos.7,301,026; 7,291,673; 7,288,553; 7,268,232; 7,268,153; 7,253,210;7,217,720; 7,214,804; 7,214,693; 7,214,690; 7,205,437; 7,186,838;7,026,500; 7,022,870; 6,998,500; 6,995,284; 6,960,474; 6,899,888;6,838,484; 6,569,896 and 6,492,554; Thevis et al., Rapid Commun MassSpectrom. 21(21): 3477-3486 (2007); Kilbourne et al., Curr Opin InvestigDrugs. 8(10): 821-829 (2007); Higuchi et al., Bioorg Med Chem Lett.17(19): 5442-5446 (2007); Zhang et al., J Med Chem. 50(16): 3857-3869(2007); Gao et al; Drug Discov Today. 12(5-6): 241-248 (2007); Omwanchaet al., Curr Opin Investig Drugs. 7(10): 873-881 (2006); Kazmin et al.,Mol Endocrinol. 20(6): 1201-1217 (2006); Segal et al., Expert OpinInvestig Drugs. 15(4): 377-387 (2006); Cadilla et al., Curr Top MedChem. 6(3): 245-270 (2006); Chen et al., Mol Interv. 5(3): 173-188(2005); Buijsman et al., Curr Med Chem. 12(9): 1017-1075 (2005); Brownet al., Endocrinology 145(12): 5417-5419 (2004); Chen et al., JPharmacol Exp Ther. 312(2): 546-553 (2005); Marhefka et al; J Med Chem.47(4): 993-998 (2004); and Yin et al., J Pharmacol Exp Ther. 304(3):1334-1340 (2003)).

This class of ligands demonstrates better pharmacokinetic andspecificity profiles than other steroidal therapies. In particular,non-steroidal SARMs display therapeutic benefit but do not display theandrogenic effects associated with other steroidal therapies. Theseadverse androgenic effects include manifestations such as prostateenlargement, acne, repression of high density lipoprotein cholesterol(HDL), hirsutism, virilization and masculinization.

The compounds provided herein are SARMs. Compounds provided hereindemonstrate the ability to exhibit their activity on the androgenreceptor in a tissue-selective manner. This tissue selectivity allowsthe compounds provided herein to function as an agonist in some tissues,while having no effect or even an antagonist effect in other tissues.The molecular basis for this tissue selective activity is not completelyunderstood. Without being limited to any particular explanation,particular ligands put nuclear receptors in different conformationalstates. These states dictate the ability of co-activators,co-repressors, and other proteins to be recruited by the nuclearreceptor. The combination of the nuclear receptor differentco-activators, co-repressors, and other proteins are the genetranscription factors that are thought to modulate tissue-selectiveeffects. Some of the SARMs exhibit agonistic anabolic properties andantagonistic androgenic properties. Compounds provided herein haveanabolic building activity, such as in muscle and bone, without unwantedandrogenic side effects on most other tissues, such as prostate and skinsebaceous glands.

Compounds provided herein are tissue-selective modulators of theandrogen receptor. In one aspect, the compounds provided herein can beused to activate the function of the androgen receptor in a subject, andin particular to activate the function of the androgen receptor in boneand/or muscle tissue and block or inhibit (antagonize) the function ofthe androgen receptor in the prostate of a male individual or in theuterus of a female individual.

In some embodiments, compounds provided herein exhibit antagonistactivity in hormone-dependent tumors while exhibiting no activity, or insome embodiments agonist activity, against other non-tumor tissuescontaining the androgen receptor. As described below, these SARMs can beused to treat a number of androgen receptor-mediated conditions,including treatment of hormone-dependent tumors containing AR, such asprostate cancer, in patients by inhibiting the growth of the tumor whilemitigating side effects such as muscle wasting, cachexia, sexualdysfunction such as loss of libido, osteoporosis and gynecomastia.

The compounds provided herein typically display micromolar orsubmicro-molar binding affinity for the androgen receptor. The compoundsprovided herein demonstrate AR agonist or antagonist activity, asevidenced by their activity in standard AR agonist and antagonistassays, such as the co-transfection assay described herein. For example,compounds provided herein demonstrate a potency (EC₅₀) of 1 μM or lessin the co-transfection assay described herein. In some embodiments,compounds provided herein demonstrate a potency (EC₅₀) of 100 nM orless. In some embodiments, compounds provided herein demonstrate apotency (EC₅₀) of 50 nM or less. In some embodiments, compounds providedherein demonstrate a potency (EC₅₀) of 10 nM or less. Compounds providedherein also demonstrate an efficacy of 50% or greater in a standard ARagonist assay. The compounds provided herein are therefore useful intreating mammals suffering from disorders related to androgen receptorfunction. Therapeutically effective amounts of one or more of thecompounds provided herein, or any of the pharmaceutically acceptablesalts thereof, are administered to the subject, to treat disordersrelated to androgen receptor function, such as, androgen deficiency,disorders that can be ameliorated by androgen replacement, or that canbe improved by androgen replacement, or to treat disorders that areresponsive to treatment with an anti-androgen. Treatment is effected byadministration of a therapeutically effective amount of one or more thanone of the compounds provided herein to a subject in need of suchtreatment. In addition, these compounds are useful as ingredients inpharmaceutical compositions alone or in combination with other activeagents.

In certain embodiments, compounds and/or compositions provided hereinare used for the prevention, treatment, or amelioration of one or moreof the symptoms of diseases or disorders associated with androgenreceptor activity. The diseases and disorders that are treated includethose caused by androgen deficiency and/or those that can be amelioratedby androgen administration, as well as those whose etiology involveshyperactivity of androgen receptor and/or those that can be amelioratedby anti-androgen administration.

Disorders, diseases or conditions that are caused by androgen deficiencyor hypoactivity or subsensitivity of androgen receptor, or that can beameliorated by androgen replacement or are responsive to treatment withan AR agonist, include, but are not limited to, aging skin; Alzheimer'sdisease; anemias, such as for example, aplastic anemia; anorexia;arthritis, including inflammatory arthritis, rheumatoid arthritis,osteoarthritis and gout; arteriosclerosis; atherosclerosis; bonedisease, including metastatic bone disease; bone damage or fracture,such as by accelerating bone fracture repair and/or stimulation ofosteoblasts and/or stimulation of bone remodeling and/or stimulation ofcartilage growth; distraction osteogenesis; reduced bone mass, densityor growth; bone weakening, such as induced by glucocorticoidadministration; musculoskeletal impairment (e.g., in the elderly);cachexia; cancer, including breast cancer and osteosarcoma; cardiacdysfunction (e.g., associated with valvular disease, myocardialinfarction, cardiac hypertrophy or congestive heart failure);cardiomyopathy; catabolic side effects of glucocorticoids; Crohn'sdisease; growth retardation in connection with Crohn's disease; shortbowel syndrome; irritable bowel syndrome; inflammatory bowel disease;ulcerative colitis; cognitive decline and impairment; dementia; shortterm memory loss; contraception (male and female); chronic obstructivepulmonary disease (COPD); chronic bronchitis; decreased pulmonaryfunction; emphysema; decreased libido in both men and women; depression;nervousness, irritability and/or stress; reduced mental energy and lowself-esteem (e.g., motivation/assertiveness); dyslipidemia; erectiledysfunction; frailty; age-related functional decline (“ARFD”) in theelderly; growth hormone deficiency; hematopoietic disorders; hormonereplacement (male and female); hyper-cholesterolemia; hyperinsulinemia;hyperlipidemia; hypertension; hyperandrogenemia; hypogonadism (includingprimary and secondary); hypothermia (including hypothermia followinganesthesia); impotence; insulin resistance; type 2 diabetes;lipodystrophy (including in subjects taking HIV or AIDS therapies suchas protease inhibitors); male menopause; metabolic syndrome (syndromeX); loss of muscle strength and/or function (e.g., in the elderly);muscular dystrophies; muscle loss following surgery (e.g., post-surgicalrehabilitation); muscular atrophy (e.g., due to physical inactivity, bedrest or reduced weight-bearing conditions such as microgravity);neurodegenerative diseases; neuromuscular disease; decreased plateletcount; platelet aggregation disorders; obesity; osteoporosis;osteopenia; glucocorticoid-induced osteoporosis;osteochondro-dysplasias; periodontal disease; premenstrual syndrome;postmenopausal symptoms in women; Reaven's syndrome; rheumatologicaldisease; sarcopenia; male and female sexual dysfunction (e.g., erectiledysfunction, decreased sex drive, sexual well-being, decreased libido);physiological short stature, including growth hormone deficient childrenand short stature associated with chronic illness and growth retardationassociated with obesity; tooth damage (such as by acceleration of toothrepair or growth); thrombocytopenia; vaginal dryness; atrophicvaginitis; ventricular dysfunction; wasting, including wasting secondaryto fractures and wasting in connection with chronic obstructivepulmonary disease (COPD), chronic liver disease, AIDS, weightlessness,cancer cachexia, burn and trauma recovery, chronic catabolic state(e.g., to coma), eating disorders (e.g., anorexia), chemotherapy,multiple sclerosis or other neurodegenerative disorders.

The compounds provided herein that demonstrate AR agonist activity alsocan be used to stimulate pulsatile growth hormone release; to improvebone strength, muscle strength and tone; to reduce subcutaneous fat in asubject; to enhance bone and muscle performance/strength; to increaseathletic performance; to attenuate or reverse protein catabolicresponses following trauma (e.g., reversal of the catabolic stateassociated with surgery, congestive heart failure, cardiac myopathy,burns, cancer, COPD); to improve sleep quality and/or correct therelative hyposomatotropism of senescence due to high increase in REMsleep and a decrease in REM latency; to treat age related decreasedtestosterone levels in men; to modifying lipid profile; and for hormonereplacement therapy, such as female androgen deficiency and maleandrogen decline.

SARMs can act as antagonists in specific tissues, and thus also areuseful in treating conditions where elevated androgen concentration oractivity causes symptoms. The compounds provided herein that demonstrateAR antagonism can be used to treat conditions whose etiology involveshyperactivity of androgen receptor or that are responsive to treatmentwith an AR antagonist. Such conditions, include, but are not limited to,acanthosis nigricans, acne, adrenal hyperandrogenism, androgeneticalopecia (male-pattern baldness), adenomas and neoplasias of theprostate (e.g., advanced metastatic prostate cancer), benign prostatehyperplasia, cancer (e.g., cancer of the breast, bladder, endometrium,lung (non-small cell lung cancer), pancreas, prostate, includingandrogen dependent prostate cancer, and skin); bulimia nervosa; chronicfatigue syndrome (CFS); chronic myalgia; acute fatigue syndrome;contraception; counteracting preeclampsia, eclampsia of pregnancy andpreterm labor; delayed wound healing; erythrocytosis; gestationaldiabetes; hirsutism; hyper-insulinemia including nesidioblastosis;hyperandrogenism; hypercortisolism; Cushing's syndrome; hyperpilosity;infertility; malignant tumor cells containing the androgen receptor,such as is the case for breast, brain, skin, ovarian, bladder,lymphatic, liver and kidney cancers; menstrual irregularity; ovarianhyperandrogenism; polycystic ovarian syndrome; seborrhea; sleepdisorders; sleep apnea; and visceral adiposity.

In some embodiments, the compounds provided herein have an agonisticactivity on muscle and bone tissue, and have a neutral or antagonisticeffect on prostate tissue. In some embodiments, such compounds are usedto treat or prevent a condition selected from among muscle wasting,cachexia, frailty, sarcopenia, osteopenia, osteoporosis, hypogonadismand sexual dysfunction.

In some embodiments, the compounds provided herein have no effect onmuscle and/or bone, and have neutral or antagonistic effect on prostate.In some embodiments, such compounds are used to treat or prevent acondition selected from among benign prostatic hyperplasia (BPH),prostate cancer and sexual dysfunction.

1. Muscle Wasting

Muscle wasting is associated with chronic, neurological, genetic orinfectious pathologies, diseases, illnesses or conditions. In someembodiments, the pathology, illness, disease or condition is chronic. Insome embodiments, the pathology, illness, disease or condition isgenetic. In some embodiments, the pathology, illness, disease orcondition is neurological. In some embodiments, the pathology, illness,disease or condition is infectious. As described herein, thepathologies, diseases, conditions or disorders for which the compoundsand compositions provided herein are administered are those thatdirectly or indirectly produce a loss of muscle mass, or that result ina muscle wasting disorder.

Muscle wasting infectious pathologies, diseases, illnesses or conditionsinclude acquired immunodeficiency syndrome (AIDS); burns; cachexias,such as AIDS cachexia, cancer cachexia, and cardiac cachexia; cancer;cardiomyopathy; chronic kidney or heart failure; chronic obstructivepulmonary disease (COPD); denervation; diabetes; emphysema; end stagerenal failure; frailty; HIV infection; inactivity; leprosy;malnutrition; muscle atrophies such as Post-Polio Muscle Atrophy (PPMA)or X-linked spinal-bulbar muscular atrophy (SBMA); muscular dystrophies,such as Duchemie Muscular Dystrophy, myotonic dystrophy, Becker'smuscular dystrophy (benign pseudohypertrophic muscular dystrophy),limb-girdle muscular dystrophy, facioscapulohumeral muscular dystrophy(FSHD), congenital muscular dystrophy, Oculopharyngeal MuscularDystrophy (OPMD), distal muscular dystrophy and Emery-Dreifuss musculardystrophy; osteomalacia; renal disease; sarcopenia; sepsis; andtuberculosis. In addition, other circumstances and conditions are linkedto and can cause muscle wasting. These include chronic lower back pain,advanced age, central nervous system (CNS) injury, peripheral nerveinjury, spinal cord injury, chemical injury, central nervous system(CNS) damage, microgravity, peripheral nerve damage, spinal cord damage,chemical damage, burns, disuse, deconditioning that occurs when a limbis immobilized, long term hospitalization due to illness or injury, andalcoholism. Muscle wasting, if left unabated, can have dire healthconsequences. For example, the changes that occur during muscle wastingcan lead to a weakened physical state that is detrimental to anindividual's health, resulting in increased susceptibility to infractionand poor performance status. In addition, muscle wasting is a strongpredictor of morbidity and mortality in patients suffering from cachexiaand AIDS.

Muscle wasting due to infectious pathologies include muscle wastingdisorders due to infection with coxsackie virus, enterovirus,Epstein-Barr virus, herpes zoster, HIV, influenza, mycobacteria,rickettsia, schistosoma, trichinella or trypanosomes.

The loss of muscle mass that occurs during muscle wasting can becharacterized by a breakdown or degradation of muscle protein, such asby muscle protein catabolism. Protein catabolism occurs because of anunusually high rate of protein degradation, an unusually low rate ofprotein synthesis, or a combination of both. Protein catabolism ordepletion, whether caused by a high degree of protein degradation or alow degree of protein synthesis, leads to a decrease in muscle mass andto muscle wasting.

Muscle wasting also is associated with advanced age. It is believed thatgeneral weakness in old age is due to muscle wasting. As the body ages,an increasing proportion of skeletal muscle is replaced by fibroustissue. The result is a significant reduction in muscle power,performance and endurance. Long term hospitalization due to illness orinjury, or muscle disuse that occurs, for example, when a limb isimmobilized, also can lead to muscle wasting. Patients sufferinginjuries, chronic illnesses, burns, trauma or cancer, who arehospitalized for long periods of time, often exhibit a long-lastingunilateral muscle wasting.

Administration of anabolic steroids have demonstrated the ability toincrease weight and muscle mass in some patients with muscle wasting,such as in cancer patients. However, administration of anabolic steroidscan result in unwanted androgenic side effects, including development ofoily skin or acne, as well as masculinization in women and prostatestimulation in men. SARMs have demonstrated efficacy for attenuatingmuscle wasting across a range of disorders (e.g., see Allen et al.Endocrine 32(1): 41-51 (2007); Lynch et al., Pharmacology & Therapeutics113(3): 461-487 (2007); Gao et al., Endocrinology 146(11): 4887-4897(2005); Lynch, Expert Opinion on Emerging Drugs 9(2): 345-361 (2004);U.S. Pat. Appl. Pub. No. 20060111441 and WO03049675). SARMs generallydemonstrate predominately anabolic activity in muscle and bone withminimal androgenic effects in most other tissues. In some embodiments,the compounds provided herein are useful for treating muscle wasting. Insome embodiments, the compounds provided herein are useful for treatingsarcopenia.

2. Muscle Tone and Strength

Androgen receptor agonists are known to have a beneficial effect onmuscle tone and strength (e.g., see Gao et al., Endocrinology 146(11):4887-4897 (2005), Jasuja et al, J Clin Endocrinol Metab. 90(2): 855-863(2005) and Ferrando et al, Am J Physiol Endocrinol Metab. 282(3):E601-E607 (2002). Androgen replacement in healthy, hypogonadal menresults in gains in fat-free mass, muscle size and maximal voluntarystrength (e.g., see Bhasin et al., J. Endocrin. 170: 27-38 (2001)).Thus, the compounds provided herein can stimulate muscle growth and canbe used for treatment of sarcopenia and frailty. In one embodiment, oneor more compounds of formula I, II or III are used to enhance muscletone in a subject. In another embodiment, one or more compounds offormula I, II or III are used to improve muscle strength in a subject.

3. Osteoporosis

Osteoporosis is a disease characterized by low bone mass and structuraldeterioration of bone tissue leading to bone fragility and an increasedsusceptibility to fractures of the hip, spine, ribs and wrist. Loss ofestrogens or androgens causes an imbalance between resorption andformation of bone by prolonging the lifespan of osteoclasts andshortening the lifespan of osteoblasts. Loss of androgens also mayinduce bone loss by increasing the rate of bone remodeling (Lindberg etal., Minerva Endocrinol. 30(1): 15-25 (2005)). The beneficial effects ofandrogens on bone in postmenopausal osteoporosis are described in art(e.g., see Hofbauer et al., Eur. J. Endocrinol. 140: 271 286 (1999).Androgens also play an important role in bone metabolism in men (e.g.,see Anderson et al., Bone 18: 171-177 (1996). Androgen receptormodulator compounds also have been shown to improve bone strength in arat model of post-menopausal osteoporosis (e.g., see Martinborough etal., J Med Chem. 50(21): 5049-5052 (2007)). Osteoporosis can result fromandrogen deprivation. In one embodiment, the compounds provided hereincan activate the function of the androgen receptor in a mammal, and inparticular to activate the function of the androgen receptor in bone.

Methods for assessing osteoporosis and osteopenia are well known in theart. For example, a subject's bone mineral density (BMD), measured bydensitometry and expressed in g/cm², is compared with a “normal value,”which is the mean BMD of sex-matched young adults at their peak bonemass, yielding a “T-score.” A score of 0 means the subject's BMD isequal to the norm for a healthy young adult. Differences between the BMDof a subject and that of the healthy young adult norm are measured instandard deviation units (SDs). A T-score between +1 and −1 isconsidered normal or healthy. A T-score between −1 and −2.5 indicateslow bone mass and is indicative of osteopenia. A T-score of −2.5 orlower is indicative of osteoporosis. As the T-score number becomes morenegative, the severity of the osteoporosis increases.

The compounds provided herein are useful for the treatment ofosteoporosis in women and men as a monotherapy or in combination withinhibitors of bone resorption, such as bisphosphonates, estrogens,SERMs, cathepsin inhibitors, α_(v)β₃ integrin receptor antagonists,calcitonin, and proton pump inhibitors. They also can be used withagents that stimulate bone formation, such as parathyroid hormone oranalogs thereof.

In one embodiment, a compound provided herein is administered incombination with an effective amount of at least one bone-strengtheningagent chosen from among estrogen and estrogen derivatives, alone or incombination with progestin or progestin derivatives; bisphosphonates;anti-estrogens; selective estrogen receptor modulators (SERMs); α_(v)β₃integrin receptor antagonists; cathepsin inhibitors; proton pumpinhibitors; PPARγ inhibitors; calcitonin; and osteoprotegerin.

Exemplary estrogen and estrogen derivatives include 17β-estradiol,estrone, conjugated estrogen (PREMARIN®), equine estrogen and17β-ethynyl estradiol. The estrogen or estrogen derivative can be usedalone or in combination with a progestin or progestin derivatives.Exemplary progestin derivatives include norethindrone andmedroxy-progesterone acetate. Other estrogen receptor modulators areknown in the art (e.g., see U.S. Pat. Nos. 7,151,196, 7,157,604,7,138,426).

Exemplary bisphosphonate compounds, which can be used in combinationwith a compound provided herein include alendronate (see U.S. Pat. Nos.5,510,517, and 5,648,491),[(cycloheptylamino)-methylene]-bis-phosphonate (incadronate) (see U.S.Pat. No. 4,970,335); (dichloromethylene)-bis-phosphonic acid (clodronicacid) and the disodium salt (clodronate) (see Quimby et al., J. Org.Chem 32: 4111-4114 (1967)); (1-hydroxy-ethylidene)-bis-phosphonate(etidronate);[1-hydroxy-3-(methylpentyl-amino)propylidene]-bis-phosphonate(ibandronate) (see U.S. Pat. No. 4,927,814),(6-amino-1-hydroxy-hexylidene)-bis-phosphonate (neridronate);[3-(dimethyl-amino)-1-hydroxy-propylidene]-bis-phosphonate(olpadronate); (3-amino-1-hydroxy-propylidene)-bis-phosphonate(pamidronate); [2-(2-pyridinyl)ethylidene]-bis-phosphonate (piridronate)(see U.S. Pat. No. 4,761,406);[1-hydroxy-2-(3-pyridinyl)-ethylidene]-bis-phosphonate (risedronate);{[(4-chlorophenyl)-thio]methylene}-bis-phosphonate (tiludronate) (seeU.S. Pat. No. 4,876,248);[1-hydroxy-2-(1H-imidazol-1-yl)-ethylidene]-bis-phosphonate(zoledronate); and[1-hydroxy-2-imidazopyridin-(1,2-a)-3-ylethylidene]-bis-phosphonate(minodronate). In one embodiment of the methods and compositionsprovided herein, the bisphosphonate is chosen from among alendronate,clodronate, etidronate, ibandronate, incadronate, minodronate,neridronate, olpadronate, pamidronate, piridronate, risedronate,tiludronate, zoledronate, and mixtures and pharmaceutically acceptablesalts thereof.

SERMs, or selective estrogen receptor modulators, are agents known inthe art to prevent bone loss by inhibiting bone resorption via pathwaysbelieved to be similar to those of estrogens (e.g., see Goldstein etal., Human Reproduction Update 6: 212-224 (2000); Lufkin et al.,Rheumatic Disease Clinics of North America 27: 163-185 (2001), Chan,Semin Oncol. 29(3 Suppl 11): 129-133 (2002), Jordan, Cancer Research 61:5683-5687 (2001) and Miller & Komm, Chapter 15, “Targeting the EstrogenReceptor with SERMs” in Ann. Rep. Med. Chem. 36: 149-158 (2001)).Exemplary SERMs include arzoxifene, clomiphene, droloxifene,enclomiphene, idoxifene, lasofoxifene, levormeloxifene, nafoxidine,raloxifene, tamoxifen, toremifene, zuclomiphene, and salts thereof (see,e.g., U.S. Pat. Nos. 4,729,999, 4,894,373 and 5,393,763).

α_(v)β₃ Integrin receptor antagonists suppress bone resorption and canbe employed in combination with the SARMs provided herein for thetreatment of bone disorders including osteoporosis. Antagonists of theα_(v)β₃ integrin receptor are known in the art (e.g., see U.S. Pat. Nos.5,204,350, 5,217,994, 5,639,754, 5,710,159, 5,723,480, 5,741,796,5,760,028, 5,773,644, 5,773,646, 5,780,426, 5,843,906, 5,852,210,5,919,792, 5,925,655, 5,929,120, 5,952,281, 5,952,341, 5,981,546,6,008,213, 6,017,925, 6,017,926, 6,028,223, 6,040,311, 6,066,648,6,069,158, 6,048,861, 6,159,964, 6,489,333, 6,784,190, 7,056,909,7,074,930 and 7,153,862.

Cathepsin is a cysteine protease and is described in U.S. Pat. Nos.5,501,969 and 5,736,357. Cysteine proteases, such as cathepsins, arelinked to a number of disease conditions, including arthritis, boneremodeling, inflammation and tumor metastasis. Cathepsin proteaseinhibitors can inhibit osteoclastic bone resorption by inhibiting thedegradation of collagen fibers and are thus useful in the treatment ofbone resorption diseases, such as osteoporosis. Examples of cathepsininhibitors are described in Deaton, Current Topics in MedicinalChemistry 5(16): 1639-1675 (2005), in U.S. Pat. Nos. 7,279,478,7,279,472, 7,112,589 and 7,012,075, and in WO 01/49288 and WO 01/77073.

Proton pump inhibitors, such as osteoclast vacuolar ATPase inhibitors,can be employed together with the SARMs provided herein. The protonATPase found on the apical membrane of the osteoclast has been reportedto play a significant role in the bone resorption process and is atarget for the design of inhibitors of bone resorption, thereby usefulfor the treatment and prevention of osteoporosis and related metabolicdiseases (e.g., see Niikura, Drug News Perspect. 19(3): 139-44 (2006),Visentin et al., J Clin Invest 106(2): 309-318 (2000) and Niikura etal., Br J of Pharmacology 142: 558-566 (2004)). Exemplary inhibitorsinclude bafilomycin A1, SB242784, FR167356, FR177995, FR202126, FR133605and NiK-12192[4-(5,6-dichloro-1H-indol-2-yl)-3-ethoxy-N-(2,2,6,6-tetramethyl-piperidin-4-yl)-benzamide](Petrangolini et al., J Pharmacol Exp Ther 318 (3): 939-946 (2006).

Activators of the peroxisome proliferator-activated receptor gamma(PPARγ) are known in the art inhibit osteoclast-like cell formation andbone resorption (e.g., see Okazaki et al., Endocrinology 140: 5060-5065(1999)). Exemplary PPARγ activators include the glitazones, such asciglitazone, darglitazone, englitazone, troglitazone, pioglitazone,rosiglitazone, and BRL 49653, the thiazolidinediones (see, e.g.,Yki-Järvinen, New Eng J Med 351(11): 1106-1118 (2004), netoglitazone,15-deoxy-Δ_(12,14)-prostaglandin J₂ and analogs, derivatives, andpharmaceutically acceptable salts thereof.

In one embodiment, therapeutically effective amounts of a compound ofFormula I, II or III alone or in combination with another active agentis administered to the mammal to treat a condition or disorder selectedfrom among osteoporosis, osteopenia, glucocorticoid-induced osteoporosisand bone fracture.

4. Prostate Disease and Prostate Cancer

The compounds provided herein can be used for treatment of prostatedisease, such as prostate cancer and benign prostatic hyperplasia (BPH).In one embodiment, compounds provided herein can block or inhibit(antagonize) the function of the androgen receptor in the prostate of amale individual.

For advanced prostate cancer, the standard treatment is androgenreceptor-blockade, usually in combination with LHRH superagonists, whichsuppresses both adrenal and testicular testosterone. The rationale ofthis approach is that early prostate cancer usually depends on androgensfor growth. The mechanism of clinically utilized anti-androgens isthought to involve blockade of the AR by binding to it and/or byinterference with binding of the AR to the DNA. Anti-androgens (androgenreceptor antagonists) have been shown to be effective for the treatmentof prostate cancer. For example, flutamide, bicalutamide and nilutamidewere found to completely block AR binding to the DNA. Bicalutamide, anon-steroidal anti-androgen, has be used to combat prostate cancer, andthe properties and usefulness of bicalutamide as an anti-androgen areknown in the art (e.g., see Fun et al., Urology 47 (Suppl. 1A): 13-25(1996) and Kolvenbag et al., Urology 47 (Suppl. 1A): 70-79 (1996)).Other examples of anti-androgens used in the treatment of prostatecancer are flutamide and nilutamide (e.g., see U.S. Pat. No. 7,018,993).The properties and usefulness of these anti-androgens are known in theart (e.g., see Neri, J. Drug Develop. 1 (Suppl.): 5-9 (1987), Neri etal., Urology 34 (4 Suppl.): 19-21 and 46-56 (1989), Neri et al., JSteroid Biochem. 6(6): 815-819 (1975), Neri et al., Anticancer Res.9(1): 13-16 (1989), Jackson et al., Anticancer Res. 27(3B): 1483-1488(2007), Harris et al., Expert Opin Investig Drugs 10(3): 493-510 (2001),Shen et al., Endocrinology 141(5): 1699-1704 (2000), Harris et al.,Drugs and Aging 3: 9-25 (1993) and U.S. Pat. No. 7,241,753). Animalmodels for determining the effectiveness of compounds for treatment ofprostate cancer are known in the art (e.g., see U.S. Pat. No.7,053,263).

In certain embodiments, compounds and/or compositions provided hereinare therapeutically effective for treating prostate cancer. In certaininstances, prostate cancer is dependant on androgens. Such androgendependent prostate cancer is typically amenable to treatment by androgenreceptor antagonists and/or androgen receptor partial agonists. Incertain embodiments, the prostate cancer is androgen dependant prostatecancer. In certain embodiments, the prostate cancer is androgenindependent prostate cancer. In certain embodiments, the prostate canceris androgen independent, but androgen receptor dependant prostatecancer.

In some embodiments, the compounds provided herein are tissue selectiveAR modulators. In some embodiments, the compounds provided herein aretissue selective AR antagonists. Tissue selective AR antagonists in theprostate that lack antagonistic action in bone and muscle are known inthe art to be useful agents for the treatment of prostate cancer, eitheralone or as an adjunct to traditional androgen deprivation therapy (see,e.g., Stoch et al., J. Clin. Endocrin. Metab. 86: 2787-2791 (2001)).Thus, compounds provided herein that are tissue selective AR antagonistsin the prostate that lack antagonistic action in bone and muscle areuseful agents for the treatment of prostate cancer, either alone or asan adjunct to traditional androgen deprivation therapy.

5. Hematopoietic Conditions and Disorders

Hematopoiesis is a constant process in which specialized blood cells,such as erythrocytes, B and T lymphocytes, platelets, granulocytes,monocytes, and macrophages, are generated from hematopoietic stem cells.A number of undesired hematopoietic conditions can occur in a subject.These include inadequate production of, or increased destruction of,platelets, red blood cells or white blood cells. For example, inadequateplatelet or blood cell production or destruction can result in aplasticanemia, refractory anemias, idiopathic thrombocytopenia purpura, immunethrombocytopenias, leukemia, myelodysplastic and preleukemia syndromes,megaloblastic anemia and platelet deficiency, myeloproliferativedisorders and uremia. Hematopoietic cytokines, such as erythropoietin,have been used to treat various diseases arising from imbalances betweendegradation and reconstitution of blood cells or from generation ofinappropriate numbers of certain blood cells.

Androgens are known in the art to stimulate renal hypertrophy anderythropoietin (EPO) production. Androgens have been used to treatanemia caused by chronic renal failure. In addition, androgens increaseserum EPO levels in anemic patients with non-severe aplastic anemia andmyelodysplastic syndromes. Thus, the selective androgen modulatorcompounds provided herein can be used to treat certain hematopoieticdisorders including aplastic anemia, refractory anemias, idiopathicthrombocytopenia purpura, immune thrombocytopenias, leukemia,preleukemia/myelodysplastic syndromes, megaloblastic anemia and plateletdeficiency, myeloproliferative disorders and uremia. In one embodiment,a compound of formula I, II or III is used to increase the number ofblood cells, such as red blood cells and platelets in a subject.

6. Neurodegenerative Diseases and Disorders

The compounds described herein can be used in the treatment ofneurodegenerative diseases, such as Alzheimer's disease. The art teachesthat androgens and selective androgen receptor modulators can be usefulin preventing the onset or delaying the progression of Alzheimer'sdisease in male patients (e.g., see Fuller et al., J Alzheimer's Dis.12(2): 129-142 (2007)). It is known in the art that androgen receptoragonists have therapeutic value in the treatment of neurodegenerativediseases such as Alzheimer's disease (e.g., see Hammond et al., J.Neurochem. 77: 1319-1326 (2001)). Androgen receptor agonists, such astestosterone, have been shown to reduce secretion of β-amyloid peptidescharacteristic of Alzheimer's disease and can therefore be used in thetreatment of Alzheimer's disease (Gouras et al., Proc. Nat. Acad. Sci.USA 97: 1202-1205 (2000)). Androgen receptor agonists also have beenshown to inhibit hyperphosphorylation of proteins implicated in theprogression Alzheimer's disease (e.g., see Papasozomenos, Proc. Nat.Acad. Sci. USA 99: 1140-1145 (2002)). Studies have shown that apoE4contributes to cognitive decline in Alzheimer's disease by reducing ARlevels in the brain, and that stimulating AR-dependent pathways canreverse apoE4-induced cognitive deficits (e.g., see Raber et al., J.Neurosci. 22(12): 5204-5209 (2002). Thus, the compounds provided hereinare useful in the treatment of Alzheimer's disease and otherneurodegenerative disorders. Additionally, androgen receptor modulatorscan be useful in treating cognitive impairment (see Pfankuch et al.,Brain Res. 1053(1-2): 88-96 (2005) and Wisniewski, Horm. Res. 58:150-155 (2002)). Studies have shown that age-related decline intestosterone levels is associated with depression and that testosteronehas been useful in the treatment of depression (e.g., see Carnahan etal., Drugs Aging 21(6): 361-376 (2004). Accordingly, the compoundsprovided herein are useful in the treatment of cognitive impairment anddepression.

7. Obesity

Obesity has been associated with alterations in androgen secretion,transport, metabolism, and action, with obese men displaying a decreaseof testosterone levels with increasing body weight and obese women,especially those with abdominal obesity, displaying a condition offunctional hyperandrogenism (e.g., see Pasquali, Fertil Steril. 85(5):1319-1340 (2006). It has been demonstrated in the art that androgenadministration reduces subcutaneous and visceral fat in obese patients(e.g., see Lovejoy et al., Int. J. Obesity 19: 614-624 (1995) andLovejoy et al., J. Clin. Endocrinol. Metab. 81: 2198-2203 (1996)).Therefore, the SARMs provided herein can be beneficial in the treatmentof obesity. In one embodiment, the compounds provided herein that are ARagonists are used to treat a male subject with abdominal adiposity. Inone embodiment, the compounds provided herein that are AR antagonistsare used to treat a female subject with abdominal obesity.

8. Insulin Disorders and Diabetes

In vivo studies have shown that the androgen receptor plays a key rolein the development of insulin resistance, which may contribute to thedevelopment of type 2 diabetes and cardiovascular disease (e.g., see Linet al., Diabetes 54(6): 1717-1725 (2005). Androgen receptor agonistsalso can have therapeutic value against metabolic syndrome (insulinresistance syndrome, syndrome X), particularly in men. Low levels oftotal and free testosterone in men have been associated with type 2diabetes, visceral obesity, insulin resistance (hyperinsulinemia,dyslipidemia) and metabolic syndrome (e.g., see Laaksonen et al.,Diabetes Care 27(5): 1036-1041 (2004), Marin et al., Obesity Res. 1(4):245-251 (1993) and Laaksonen et al., Euro. J Endocrin 149: 601-608(2003)) and, in women, there is a correlation between high androgenlevels and insulin resistance (e.g., see Corbould, J Endocrinol. 192(3):585-594 (2007). Accordingly, the compounds provided herein can be usedto treat insulin resistance and type II diabetes.

9. Sexual Dysfunction

Testosterone is used as a treatment for sexual dysfunction inhypogonadal patients (Yassin et al., World Journal of Urology 24:6:639(2006). It is known in the art that androgen deficiency in women isclinically often associated with a loss of libido and energy (e.g.,Arlt, Eur J Endocrinol 154(1): 1-11 (2006) and Rivera-Woll et al., HumanReproduction Update 10(5): 421 (2004)). Low androgen levels have beenshown to contribute to the decline in sexual interest in many womenduring their later reproductive years (Davis, Clin. Endocrinol. Metab.84: 1886-1891 (1999)). In clinical trials, women treated with theandrogen DHEA exhibited an increase in the frequency of sexual thoughts,interest, and satisfaction compared to women taking a placebo (e.g., seeArlt et al., N Engl. J. Med. 341: 1013-1020 (1999) and Miller, J. Clin.Endocrinol. Metab. 86: 2395-2401 (2001)). Androgen deficiency in men isrelated to diminished libido (e.g., see Fine, JAOA Supplement 1 Vol104(1): S9-S15 (2004)). It also is known in the art that erectileresponse is centrally and peripherally regulated by androgens. Studieshave shown that treatment with testosterone positively impacts thetissues of the penis involved in the mechanism of erection, and thattestosterone deficiency impairs the anatomical and physiologicalerectile capacity, which is reversible upon androgen replacement (e.g.,see Gooren et al., Asian Journal of Andrology 8(1): 3-9 (2006). Thus,the compounds provided herein can be useful in the treatment of sexualdysfunction. For example, the compounds provided herein are useful forhormone replacement therapy in hypogonadic (androgen deficient) men.

In one embodiment, compounds provided herein are useful in activatingthe function of the androgen receptor in bone and/or muscle tissue andblocking or inhibiting the function of the androgen receptor in theprostate of a male individual or in the uterus of a female individual.

In one embodiment, the compounds provided herein are used to attenuateor block the function of the androgen receptor in the prostate of a maleindividual or in the uterus of a female individual induced by ARagonists without effecting hair-growing skin or vocal cords. In oneembodiment, the compounds provided herein are used to activate thefunction of the androgen receptor in bone and/or muscle tissue, but notin organs which control blood lipid levels (e.g., the liver).

10. Arthritic Conditions and Inflammatory Disorders

Androgen receptor modulators are known in the art to be useful in thetreatment of arthritic conditions or inflammatory disorders (e.g., seeCutolo et al., Ann. N.Y. Acad. Sci. 966: 131-142 (2002); Cutolo, RheumDis Clin North Am 26(4): 881-895 (2000); Bijlsma et al., Am J ReprodImmunol 28(34): 231-234 (1992); Jansson et al., Arthritis Rheum 44(9):2168-2175 (2001); and Purdie, Br Med Bull 56(3): 809-823 (2000). Also,see Merck Manual, 17th edition, pp. 449-451.)

The compounds provided herein also are used, alone or in combination, totreat or prevent arthritic conditions, such as Behcet's disease,bursitis, tendonitis, CPPD deposition disease, carpal tunnel syndrome,Ehlers-Danlos syndrome, fibromyalgia, gout, infectious arthritis,inflammatory bowel disease, juvenile arthritis, lupus erythematosus,Lyme disease, Marfan syndrome, myositis, osteoarthritis, osteogenesisimperfecta, osteonecrosis, polyarteritis, polymyalgia rheumatica,psoriatic arthritis, Raynaud's phenomenon, reflex sympathetic dystrophysyndrome, Reiter's syndrome, rheumatoid arthritis, scleroderma andSjogren's syndrome. Provided herein are methods for the treatment orprevention of an arthritic condition, the method including administeringa therapeutically effective amount of a compound of any of Formulae I,II or III in an amount effective for the treatment or prevention of anarthritic condition or an inflammatory disorder. In one embodiment, themethod is for the treatment or prevention of osteoarthritis, whichincludes administering a therapeutically effective amount of a compoundof any of Formulae I, II or III in an amount effective for the treatmentor prevention of osteoarthritis.

11. Modifying Lipid Profile

High doses of testosterone and other anabolic steroids have the abilityto reduce cholesterol and to reduce HDL, often greater that 60%, 65%,70%, 75% and 80%. In one embodiment, a compound of formula I, II or IIIis used to reduce total cholesterol, LDL, HDL, VLDL, and/ortriglycerides. In some embodiments, administration of a compound offormula I, II or III can be used to reduce levels of total cholesterol,LDL, VLDL, triglycerides and/or HDL, while the LDL/HDL ratio remains inthe normal range.

12. Contraception

Hormonal contraception is known in the art. For example, hormonal malecontraceptive methods provide pregnancy protection by means ofspermatogenic suppression or inhibition, generally by the suppression ofgonadotropins (e.g., see Pasqualotto et al., Rev Hosp Clin Fac Med SaoPaulo 58(5): 275-283 (2003), Ly et al., Hum Reprod. 20(6): 1733-1740(2005) and Brady et al., Hum Reprod. 19(11): 2658-2667 (2004)). This canbe accomplished by administration of an androgen receptor modulatoralone, or in combination with other androgens, such as19-nortestosterone, 7α-methyl-19-nortestosterone (MENT) and5α-dihydrotestosterone, or administration of testosterone in combinationwith an anti-androgen, (e.g., see Turner et al., J Clin Endocrinol Metab88(10): 4659-4667 (2003). Hormonally mediated suppression of ovulationin women using synthetic androgens is known in the art, but their use islimited because of adverse effects, including weight gain, decreasedhigh-density lipoprotein cholesterol, and in some instances, facial hairgrowth and acne (e.g., see Johnson, Clin Obstet Gynecol. 41: 405-421(1998) and Sulak, Am J Manag Care 11: S492-S497 (2005). The selectiveandrogen modulating compounds provided herein are useful for providingcontraception while minimizing adverse side effects associated withsteroidal androgens.

13. Postmenopausal Conditions

Reduced levels of testosterone in postmenopausal women are associatedwith loss of libido, decreased sexual activity, diminished feelings ofphysical well-being and fatigue (e.g., see Kingsberg, J Sex Med. 4 Suppl3: 227-234 (2007). The compounds disclosed herein can exhibit androgenagonism in the central nervous system and can be used to treat vasomotorsymptoms, such as hot flashes, and other postmenopausal conditions, andto increase energy. There is evidence in the art that hot flashesdecrease in women treated with androgens (e.g., see Notelovitz, MayoClin Proc. 79(4 Suppl): S8-S 13 (2004)). Tissue-selective AR antagonistsalso can treat polycystic ovarian syndrome in postmenopausal women(Eagleson et al., J. Clin. Endocrinol. Metab. 85: 4047-4052 (2000)). Inone embodiment, a compound of formula I, II or III is used to treat apostmenopausal condition in a female subject. In one embodiment, thepostmenopausal condition is selected from among hot flashes, loss oflibido, decreased feelings of well being and fatigue. In one embodiment,a compound of formula I, II or III is used to treat hot flashes in afemale subject. In one embodiment, a compound of formula I, II or III isused to treat hypoactive sexual desire disorder in a postmenopausalfemale subject.

E. FORMULATION OF PHARMACEUTICAL COMPOSITIONS

The compounds of Formula I, II or III or pharmaceutically acceptablesalts or prodrugs thereof can be provided as combinations with othertherapeutic agents or in pharmaceutical compositions. The pharmaceuticalcompositions provided herein include therapeutically effective amountsof one or more of the selective androgen receptor activity modulatorsprovided herein that are useful in the prevention, treatment, oramelioration of one or more of the symptoms of diseases or disordersassociated with androgen receptor activity.

The compositions include one or more compounds provided herein. Thecompositions are formulated into suitable pharmaceutical preparationssuch as solutions, suspensions, tablets, dispersible tablets, pills,capsules, powders, sustained release formulations or elixirs, for oraladministration or in sterile solutions or suspensions for parenteraladministration, as well as transdermal patch preparation and dry powderinhalers. Typically the compounds described above are formulated intopharmaceutical compositions using techniques and procedures well knownin the art (see, e.g., Ansel, Introduction to Pharmaceutical DosageForms, 4^(th) Edition (1985), 126).

In certain embodiments, a pharmaceutical composition including one ormore compounds provided herein is prepared using known techniques,including, but not limited to, mixing, dissolving, granulating,dragée-making, levigating, emulsifying, encapsulating, entrapping ortabletting processes.

In the compositions, effective concentrations of one or more compoundsor pharmaceutically acceptable derivatives is (are) mixed with asuitable pharmaceutical carrier or vehicle. The compounds can bederivatized as the corresponding salts, esters, enol ethers or esters,acids, bases, solvates, hydrates or prodrugs prior to formulation, asdescribed above. The concentrations of the compounds in the compositionsare effective for delivery of an amount, upon administration, thattreats, prevents, or ameliorates one or more of the symptoms of diseasesor disorders associated with androgen activity or in which androgenactivity is implicated.

Typically, the compositions are formulated for single dosageadministration. To formulate a composition, the weight fraction ofcompound is dissolved, suspended, dispersed or otherwise mixed in aselected vehicle at an effective concentration such that the treatedcondition is relieved or ameliorated. Pharmaceutical carriers orvehicles suitable for administration of the compounds provided hereininclude any such carriers known to those skilled in the art to besuitable for the particular mode of administration.

In addition, the compounds can be formulated as the solepharmaceutically active ingredient in the composition or can be combinedwith other active ingredients. Liposomal suspensions, includingtissue-targeted liposomes, such as tumor-targeted liposomes, also can besuitable as pharmaceutically acceptable carriers. These can be preparedaccording to methods known to those skilled in the art. For example,liposome formulations can be prepared as described in U.S. Pat. No.4,522,811. Briefly, liposomes such as multilamellar vesicles (MLV's) canbe formed by drying down egg phosphatidyl choline and brain phosphatidylserine (7:3 molar ratio) on the inside of a flask. A solution of acompound provided herein in phosphate buffered saline lacking divalentcations (PBS) is added and the flask shaken until the lipid film isdispersed. The resulting vesicles are washed to remove unencapsulatedcompound, pelleted by centrifugation, and then resuspended in PBS.

One or more than one of the compounds provided herein is included in thepharmaceutically acceptable carrier in an amount sufficient to exert atherapeutically useful effect in the absence of undesirable side effectson the subject treated.

The concentration of the one or more than one compounds provided hereinin the pharmaceutical composition will depend on absorption,inactivation and excretion rates of the compound, the physicochemicalcharacteristics of the compound, the dosage schedule, and amountadministered as well as other factors known to those of skill in theart. For example, the amount that is delivered is sufficient toameliorate one or more of the symptoms of diseases or disordersassociated with androgen activity or in which androgen activity isimplicated, as described herein.

The effective amount of a compound provided herein can be determined byone of ordinary skill in the art, and includes exemplary dosage amountsfor a mammal of from about 0.05 to 100 mg/kg of body weight of activecompound per day, which can be administered in a single dose or in theform of individual divided doses, such as from 1 to 4 times per day. Itwill be understood that the specific dose level and frequency of dosagefor any particular subject can be varied and will depend upon a varietyof factors, including the activity of the specific compound employed,the metabolic stability and length of action of that compound, thespecies, age, body weight, general health, sex and diet of the subject,the mode and time of administration, rate of excretion, drugcombination, and severity of the particular condition. In someembodiments, the daily dosage of a compound provided herein can bevaried over a wide range from about or 0.01 to about or 1000 mg peradult human per day. For example, dosages can range from about or 0.1 toabout or 200 mg/day. In some embodiments, the dosage can range from 0.2mg to 20 mg per day. In some embodiments, the dosage can range from 0.5mg to 10 mg per day. In some embodiments, the daily dosage can be 0.1mg, 0.25 mg, 0.5 mg, 0.75 mg, 1 mg, 1.25 mg, 1.5 mg, 1.75 mg, 2 mg, 2.25mg, 2.5 mg, 2.75 mg, 3 mg, 3.25 mg, 3.5 mg, 3.75 mg, 4 mg, 4.25 mg, 4.5mg, 4.75 mg, 5 mg, 5.25 mg, 5.5 mg, 5.75 mg, 6 mg, 6.25 mg, 6.5 mg, 6.75mg, 7 mg, 7.25 mg, 7.5 mg, 7.75 mg, 8 mg, 8.25 mg, 8.5 mg, 8.75 mg, 9mg, 9.25 mg, 9.5 mg, 9.75 mg, 10 mg. For oral administration, thecompositions can be provided in the form of unit dosages such as tabletsor capsules or liquids including from about or 0.01 to about or 1000 mg,such as for example, 0.01, 0.05, 0.075, 0.1, 0.25, 0.5, 0.75, 1, 2, 2.5,3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 50, 75, 100, 125, 150, 175, 180,190, 200, 225, 250, 300, 400, 500, 750, 800, 850, 900, 950 and 1000milligrams of the active ingredient for the symptomatic adjustment ofthe dosage to the subject to be treated. In some embodiments, thecompositions can be provided in the form of unit dosages such as tabletsor capsules or liquids including from about or 0.01 to about or 1000 μg,such as for example, 0.01, 0.05, 0.075, 0.1, 0.25, 0.5, 0.75, 1, 2.5, 3,4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 50, 75, 100, 125, 150, 175, 180, 190,200, 225, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800,850, 900, 950 and 1000 micrograms of the active ingredient for thesymptomatic adjustment of the dosage to the subject to be treated.

The pharmaceutical composition including one or more than one compoundprovided herein can be administered at once, or can be divided into anumber of smaller doses to be administered at intervals of time. It isunderstood that the precise dosage and duration of treatment is afunction of the disease being treated and can be determined empiricallyusing known testing protocols or by extrapolation from in vivo or invitro test data. It is to be noted that concentrations and dosage valuesalso can vary with the severity of the condition to be alleviated. It isto be further understood that for any particular subject, specificdosage regimens should be adjusted over time according to the individualneed and the professional judgment of the person administering orsupervising the administration of the compositions, and that theconcentration ranges set forth herein are exemplary only and are notintended to limit the scope or practice of the compounds, compositions,methods and other subject matter provided herein.

Pharmaceutically acceptable derivatives include acids, bases, enolethers and esters, salts, esters, hydrates, solvates and prodrug forms.The derivative is selected such that its pharmacokinetic properties aresuperior to the corresponding neutral compound.

Thus, effective concentrations or amounts of one or more of thecompounds described herein or pharmaceutically acceptable derivatives orprodrugs thereof are mixed with a suitable pharmaceutical carrier orvehicle for systemic, topical or local administration to formpharmaceutical compositions. One or more than one compound providedherein is/are included in an amount effective for ameliorating one ormore symptoms of, or for treating or preventing diseases or disordersassociated with androgen receptor activity or in which androgen receptoractivity is implicated, as described herein. The concentration of theone or more than one compound in the composition will depend onabsorption, inactivation, excretion rates of the active compound, thedosage schedule, amount administered, particular formulation as well asother factors known to those of skill in the art.

The compositions are intended to be administered by a suitable route,including orally in the form of capsules, tablets, granules, powders orliquid formulations including syrups; parenterally, such assubcutaneously, intravenously, intramuscularly, with intersternalinjection or infusion techniques (as sterile injectable aqueous (aq.) ornon-aqueous solutions or suspensions); nasally, such as by inhalationspray; topically, such as in the form of a cream or ointment; rectally,such as in the form of suppositories; liposomally; and locally. Thecompositions can be in liquid, semi-liquid or solid form and areformulated in a manner suitable for each route of administration. Incertain embodiments, administration of the formulation includesparenteral and oral modes of administration. In one embodiment, thecompositions are administered orally.

In certain embodiments, the pharmaceutical compositions provided hereinincluding one or more compounds provided herein is a solid (e.g., apowder, tablet, and/or capsule). In certain of such embodiments, a solidpharmaceutical composition including one or more compounds providedherein is prepared using ingredients known in the art, including, butnot limited to, starches, sugars, diluents, granulating agents, gums,lubricants, binders, and disintegrating agents.

In certain embodiments, a pharmaceutical composition including one ormore compounds provided herein is formulated as a depot preparation.Certain of such depot preparations are typically longer acting thannon-depot preparations. In certain embodiments, such preparations areadministered by implantation (for example, subcutaneously orintramuscularly) or by intramuscular injection. In certain embodiments,depot preparations are prepared using suitable polymeric or hydrophobicmaterials (for example, an emulsion in an acceptable oil) or ionexchange resins, or as sparingly soluble derivatives, for example, as asparingly soluble salt.

In certain embodiments, a pharmaceutical composition including one ormore compounds provided herein includes a delivery system. Examples ofdelivery systems include, but are not limited to, liposomes andemulsions. Certain delivery systems are useful for preparing certainpharmaceutical compositions including those including hydrophobiccompounds. In certain embodiments, certain organic solvents such asdimethylsulfoxide are used.

In certain embodiments, a pharmaceutical composition including one ormore compounds provided herein (active ingredient) includes one or moretissue-specific delivery molecules designed to deliver thepharmaceutical composition to specific tissues or cell types. Forexample, in certain embodiments, pharmaceutical compositions includeliposomes coated with a tissue-specific antibody.

In certain embodiments, a pharmaceutical composition including one ormore compounds provided herein includes a co-solvent system. Certain ofsuch co-solvent systems include, for example, benzyl alcohol, a nonpolarsurfactant, a water-miscible organic polymer, and an aqueous phase. Incertain embodiments, such co-solvent systems are used for hydrophobiccompounds. A non-limiting example of such a co-solvent system is the VPDco-solvent system, which is a solution of absolute ethanol including 3%w/v benzyl alcohol, 8% w/v of the nonpolar surfactant Polysorbate 80™,and 65% w/v polyethylene glycol 300. The proportions of such co-solventsystems can be varied considerably without significantly altering theirsolubility and toxicity characteristics. Furthermore, the identity ofco-solvent components can be varied: for example, other surfactants canbe used instead of Polysorbate 80™; the fraction size of polyethyleneglycol can be varied; other biocompatible polymers can replacepolyethylene glycol, e.g., polyvinyl pyrrolidone; and other sugars orpolysaccharides can substitute for dextrose.

In certain embodiments, solutions or suspensions used for parenteral,intradermal, subcutaneous, or topical application can include any of thefollowing components: a sterile diluent, such as water for injection,saline solution, fixed oil, polyethylene glycol, glycerine, propyleneglycol or other synthetic solvent; antimicrobial agents, such as benzylalcohol and methyl parabens; antioxidants, such as ascorbic acid andsodium bisulfate; chelating agents, such as ethylenediamine-tetraaceticacid (EDTA); buffers, such as acetates, citrates and phosphates; andagents for the adjustment of tonicity such as sodium chloride ordextrose. Parenteral preparations can be enclosed in ampules, disposablesyringes or single or multiple dose vials made of glass, plastic orother suitable material.

In instances in which the compounds exhibit insufficient solubility,methods for solubilizing compounds can be used. Such methods are knownto those of skill in this art, and include, but are not limited to,using cosolvents, such as dimethylsulfoxide (DMSO), using surfactants,such as surfactants that include polyoxyethylene derivatives of sorbitanmonolaurate, such as TWEEN® or polysorbate surfactants, or dissolutionin aqueous sodium bicarbonate. Derivatives of the compounds, such asprodrugs of the compounds also can be used in formulating effectivepharmaceutical compositions.

In certain embodiments, a pharmaceutical composition including one ormore compounds provided herein includes a sustained release system. Anon-limiting example of such a sustained-release system is asemipermeable matrix of solid hydrophobic polymers. In certainembodiments, sustained release systems can, depending on their chemicalnature, release compounds over a period of hours, days, weeks or months.

In certain embodiments, upon mixing or addition of the compound(s), theresulting mixture can be a solution, suspension or emulsion. The form ofthe resulting mixture depends upon a number of factors, including theintended mode of administration and the solubility of the compound inthe selected carrier or vehicle. The effective concentration issufficient for ameliorating the symptoms of the disease, disorder orcondition treated and can be empirically determined.

The pharmaceutical compositions are provided for administration tohumans and animals in unit dosage forms, such as tablets, capsules,pills, powders, granules, sterile parenteral solutions or suspensions,and oral solutions or suspensions, and oil-water emulsions includingsuitable quantities of the compounds or pharmaceutically acceptablederivatives thereof. The pharmaceutically active compounds andderivatives thereof are typically formulated and administered in unitdosage forms.

The composition can include in addition to the one or more than onecompound provided herein other ingredients, such as, but not limited to,a diluent such as lactose, sucrose, dicalcium phosphate, orcarboxymethylcellulose; a lubricant, such as magnesium stearate, calciumstearate and talc; and a binder such as starch, natural gums, such asgum acacia, gelatin, glucose, molasses, polyvinylpyrrolidone, cellulosesand derivatives thereof, povidone, crospovidones and other such bindersknown to those of skill in the art. Liquid pharmaceuticallyadministrable compositions can, for example, be prepared by dissolving,dispersing, or otherwise mixing an active compound as defined above andoptional pharmaceutical adjuvants in a carrier, such as, for example,water, saline, aqueous dextrose, glycerol, glycols and ethanol, tothereby form a solution or suspension. If desired, the pharmaceuticalcomposition to be administered also can include minor amounts ofnontoxic auxiliary substances such as wetting agents, emulsifyingagents, or solubilizing agents, or pH buffering agents, for example,acetate or sodium citrate, or cyclodextrin derivatives, sorbitanmonolaurate, triethanolamine sodium acetate, triethanolamine oleate.Actual methods of preparing such dosage forms are known, or will beapparent, to those skilled in this art; for example, see Remington'sPharmaceutical Sciences, Mack Publishing Company, Easton, Pa., 15^(th)edition (1975). The composition or formulation to be administered will,in any event, include a quantity of the active compound in an amountsufficient to alleviate the symptoms of the treated subject.

Dosage forms or compositions can be prepared to include one or more thanone compound provided herein in the range of 0.005% to 100% with thebalance made up from non-toxic carrier. For oral administration, apharmaceutically acceptable non-toxic composition is formed by theincorporation of any of the normally employed excipients, such as, forexample pharmaceutical grades of mannitol, lactose, starch, magnesiumstearate, talcum, cellulose derivatives, sodium croscarmellose, glucose,sucrose, magnesium carbonate or sodium saccharin. Such compositionsinclude solutions, suspensions, tablets, capsules, powders and sustainedrelease formulations, such as, but not limited to, implants andmicroencapsulated delivery systems, and biodegradable, biocompatiblepolymers, such as collagen, ethylene vinyl acetate, polyanhydrides,polyglycolic acid, polyorthoesters, polylactic acid and others. Methodsfor preparation of these compositions are known to those skilled in theart. The contemplated compositions can include 0.001%-100% activeingredient, in one embodiment 0.1-85%, in another embodiment 75-95%. Insome embodiments, the compositions include 1-10% active ingredient. Insome embodiments, the compositions include 10-25% active ingredient. Insome embodiments, the compositions includes 15-35% active ingredient. Insome embodiments, the compositions include 40-60% active ingredient. Insome embodiments, the compositions include 50-75% active ingredient. Insome embodiments, the active ingredient is present at 1%, 2%, 3%, 4%,5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%,20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%,34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%,48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%,62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%,76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%,90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%.

In certain embodiments, the compounds can be administered in a formsuitable for immediate release or extended release. Immediate release orextended release can be achieved with suitable pharmaceuticalcompositions or, particularly in the case of extended release, withdevices such as subcutaneous implants or osmotic pumps. Exemplarycompositions for topical administration include a topical carrier suchas a mineral oil gelled with polyethylene (e.g., PLASTIBASE®).

In certain embodiments, compounds provided herein used in thepharmaceutical compositions can be provided as pharmaceuticallyacceptable salts with pharmaceutically compatible counter-ions.Pharmaceutically compatible salts can be formed with many acids,including, but not limited to, hydrochloric, sulfuric, acetic, citric,ascorbic, butyric, lactic, tartaric, malic, fumaric, succinic andvaleric.

In certain embodiments, the pharmaceutical compositions include one ormore than one compound provided herein in a therapeutically effectiveamount. In certain embodiments, the therapeutically effective amount issufficient to prevent, alleviate or ameliorate symptoms of a disease orto prolong the survival of the subject being treated. Determination of atherapeutically effective amount is well within the capability of thoseskilled in the art.

The compositions can include in addition to the one or more than onecompound provided herein other active compounds to obtain desiredcombinations of properties. The compounds provided herein, orpharmaceutically acceptable derivatives or prodrugs thereof as describedherein, also can be advantageously administered for therapeutic orprophylactic purposes together with another pharmacological agent knownin the general art to be of value in treating one or more of thediseases or medical conditions referred to hereinabove, such as diseasesor disorders associated with androgen receptor activity or in whichandrogen receptor activity is implicated. It is to be understood thatsuch combination therapy constitutes a further aspect of thecompositions and methods of treatment provided herein.

In certain embodiments, a pharmaceutical composition including one ormore compounds provided herein is formulated as a prodrug. In certainembodiments, prodrugs are useful because they are easier to administerthan the corresponding active form. For example, in certain instances, aprodrug can be more bioavailable (e.g., through oral administration)than is the corresponding active form. In certain instances, a prodrugcan have improved solubility compared to the corresponding active form.In certain embodiments, a prodrug is an ester. In certain embodiments,such prodrugs are less water soluble than the corresponding active form.In certain instances, such prodrugs possess superior transmittal acrosscell membranes, where water solubility is detrimental to mobility. Incertain embodiments, the ester in such prodrugs is metabolicallyhydrolyzed to carboxylic acid. In certain embodiments, a prodrugincludes a short peptide (polyamino acid) bound to an acid group. Incertain of such embodiments, the peptide is metabolized to form thecorresponding active form.

In certain embodiments, a pharmaceutical composition including one ormore compounds provided herein is useful for treating a conditions ordisorder in a mammalian, and particularly in a human subject. Suitableadministration routes include, but are not limited to, oral, rectal,transmucosal, intestinal, enteral, topical, suppository, throughinhalation, intrathecal, intraventricular, intraperitoneal, intranasal,intraocular and parenteral (e.g., intravenous, intramuscular,intramedullary, and subcutaneous). In certain embodiments,pharmaceutical compositions are administered to achieve local ratherthan systemic exposures. For example, pharmaceutical compositions can beinjected directly in the area of desired effect (e.g., in the renal orcardiac area). In certain embodiments in which the pharmaceuticalcomposition is administered locally, the dosage regimen is adjusted toachieve a desired local concentration of a compound provided herein.

In certain embodiments, a pharmaceutical composition including one ormore compounds provided herein is administered in the form of a dosageunit (e.g., tablet, capsule, pill, injection, bolus). In certainembodiments, such dosage units include a selective androgen receptormodulator provided herein in a dose from about or 0.01 μg/kg of bodyweight to about or 50 mg/kg of body weight. In certain embodiments, suchdosage units include a selective androgen receptor modulator in a dosefrom about or 0.05 μg/kg of body weight to about or 40 mg/kg of bodyweight. In certain embodiments, such dosage units include a selectiveandrogen receptor modulator in a dose from about or 0.1 μg/kg of bodyweight to about or 30 mg/kg of body weight. In certain embodiments, suchdosage units include a selective androgen receptor modulator in a dosefrom about or 0.5 μg/kg of body weight to about or 25 mg/kg of bodyweight. In certain embodiments, such dosage units include a selectiveandrogen receptor modulator in a dose from about or 1 μg/kg of bodyweight to about or 20 mg/kg of body weight. In certain embodiments, suchdosage units include a selective androgen receptor modulator in a dosefrom about or 2 μg/kg of body weight to about or 15 mg/kg of bodyweight. In certain embodiments, such dosage units include a selectiveandrogen receptor modulator in a dose from about or 10 μg/kg of bodyweight to about or 5 mg/kg of body weight. In certain embodiments, suchdosage units include a selective androgen receptor modulator providedherein in a dose from about or 0.01 mg/kg of body weight to about or 1mg/kg of body weight. In certain embodiments, such dosage units includea selective androgen receptor modulator in a dose from about or 0.05mg/kg of body weight to about or 0.1 mg/kg of body weight. In certainembodiments, such dosage units include a selective androgen receptormodulator in a dose from about or 0.001 μg/kg of body weight to about or100 μg/kg of body weight. In certain embodiments, such dosage unitsinclude a selective androgen receptor modulator in a dose from about or0.01 μg/kg of body weight to about or 10 μg/kg of body weight. Incertain embodiments, such dosage units include a selective androgenreceptor modulator in a dose from about or 0.1 μg/kg of body weight toabout or 1 μg/kg of body weight. An approximate average adult bodyweight is 70 kg. Thus, for an adult of average body weight, a dose of0.1 μg/kg of body weight is equivalent to 7 μg, a dose of 1 μg/kg ofbody weight is equivalent to 70 μg, a dose of 10 μg/kg of body weight isequivalent to 700 μg or 0.7 mg and a dose of 0.1 mg/kg of body weight isequivalent to 7 mg.

In certain embodiments, pharmaceutical compositions are administered asneeded, once per day, twice per day, three times per day, or four ormore times per day. It is recognized by those skilled in the art thatthe particular dose, frequency, and duration of administration dependson a number of factors, including, without limitation, the biologicalactivity desired, the condition of the subject, and tolerance for thepharmaceutical composition.

In certain embodiments, a pharmaceutical composition provided herein isadministered for a period of continuous therapy. For example, apharmaceutical composition provided herein can be administered over aperiod of days, weeks, months, or years.

Dosage amount, interval between doses, and duration of treatment can beadjusted to achieve a desired effect. In certain embodiments, dosageamount and interval between doses are adjusted to maintain a desiredconcentration of compound in a subject. For example, in certainembodiments, dosage amount and interval between doses are adjusted toprovide plasma concentration of a compound provided herein at an amountsufficient to achieve a desired effect. In certain of such embodimentsthe plasma concentration is maintained above the minimal effectiveconcentration (MEC). In certain embodiments, pharmaceutical compositionsprovided herein are administered with a dosage regimen designed tomaintain a concentration above the MEC for 10-90% of the time, between30-90% of the time, or between 50-90% of the time.

1. Compositions for Oral Administration

In certain embodiments, oral pharmaceutical dosage forms are eithersolid, gel or liquid. The solid dosage forms are tablets, capsules,granules, and bulk powders. Types of oral tablets include compressed,chewable lozenges and tablets which can be enteric coated, sugar coatedor film coated. Capsules can be hard or soft gelatin capsules, whilegranules and powders can be provided in non-effervescent or effervescentform with the combination of other ingredients known to those skilled inthe art.

In certain embodiments, the formulations are solid dosage forms, such ascapsules or tablets. The tablets, pills, capsules, troches and othersolid dosage forms can include any of the following ingredients, orcompounds of a similar nature: a binder; a diluent; a disintegratingagent; a lubricant; a glidant; a sweetening agent; and a flavoringagent.

In certain embodiments, pharmaceutical compositions for oraladministration are push fit capsules made of gelatin. Certain of suchpush fit capsules include one or more compounds provided herein inadmixture with one or more fillers such as lactose, binders such asstarches, and/or lubricants such as talc or magnesium stearate and,optionally, stabilizers. In certain embodiments, pharmaceuticalcompositions for oral administration are soft, sealed capsules made ofgelatin and a plasticizer, such as glycerol or sorbitol. In certain softcapsules, one or more compounds provided are be dissolved or suspendedin suitable liquids, such as fatty oils, liquid paraffin, or liquidpolyethylene glycols. In addition, stabilizers can be added.

In certain embodiments, pharmaceutical compositions are prepared forbuccal administration. Certain of such pharmaceutical compositions aretablets or lozenges formulated in conventional manner. In someembodiments, the compositions are formulated as dissolvable films, suchas those made with pullulan or described in the art (e.g., see U.S. Pat.Nos. 6,596,298, 7,067,116, 7,182,964 and 7,241,411).

Examples of binders for use in the compositions provided herein includemicrocrystalline cellulose, gum tragacanth, glucose solution, gumarabic, gelatin solution, sucrose and starch paste. Lubricants includetalc, starch, magnesium or calcium stearate, lycopodium and stearicacid. Diluents include, for example, lactose, sucrose, starch, kaolin,salt, mannitol and dicalcium phosphate. Glidants include, but are notlimited to, colloidal silicon dioxide. Disintegrating agents includecroscarmellose sodium, sodium starch glycolate, alginic acid, sodiumalginate, corn starch, potato starch, bentonite, methylcellulose, agarand carboxymethylcellulose. Coloring agents include, for example, any ofthe approved certified water soluble FD and C dyes, mixtures thereof;and water insoluble FD and C dyes suspended on alumina hydrate.Sweetening agents include sucrose, lactose, mannitol, xylitol andartificial sweetening agents such as saccharin. Flavoring agents includenatural flavors extracted from plants such as fruits and syntheticblends of compounds which produce a pleasant sensation, such as, but notlimited to peppermint and methyl salicylate, including spray driednatural and artificial flavors. Wetting agents include propylene glycolmonostearate, sorbitan monooleate, diethylene glycol monolaurate andpolyoxyethylene laural ether. Emetic coatings include fatty acids, fats,waxes, shellac, ammoniated shellac and cellulose acetate phthalates.Film coatings include hydroxyethylcellulose, sodiumcarboxymethylcellulose, polyethylene glycol 4000 and cellulose acetatephthalate.

If oral administration is desired, the compound can be provided in acomposition that protects it from the acidic environment of the stomach.For example, the composition can be formulated in an enteric coatingthat maintains its integrity in the stomach and releases the activecompound in the intestine. The composition also can be formulated incombination with an antacid or other such ingredient.

When the dosage unit form is a capsule, it can include, in addition tomaterial of the above type, a liquid carrier such as a fatty oil. Inaddition, dosage unit forms can include various other materials whichmodify the physical form of the dosage unit, for example, coatings ofsugar and other enteric agents. The compounds also can be administeredas a component of an elixir, suspension, syrup, wafer, sprinkle orchewing gum. A syrup can include, in addition to the active compounds,sucrose as a sweetening agent and certain preservatives, dyes andcolorings and flavors.

The active materials also can be mixed with other active materials whichdo not impair the desired action, or with materials that supplement thedesired action, such as antacids, H2 blockers (acid reducers), anddiuretics.

Pharmaceutically acceptable carriers included in tablets are binders,lubricants, diluents, disintegrating agents, coloring agents, flavoringagents, and wetting agents. Enteric coated tablets, because of theenteric coating, resist the action of stomach acid and dissolve ordisintegrate in the neutral or alkaline intestines. Sugar coated tabletsare compressed tablets to which different layers of pharmaceuticallyacceptable substances are applied. Film coated tablets are compressedtablets which have been coated with a polymer or other suitable coating.Multiple compressed tablets are compressed tablets made by more than onecompression cycle utilizing the pharmaceutically acceptable substancespreviously mentioned. Coloring agents also can be used in the abovedosage forms. Flavoring and sweetening agents are used in compressedtablets, sugar coated, multiple compressed and chewable tablets.Flavoring and sweetening agents are especially useful in the formationof chewable tablets and lozenges.

Liquid oral dosage forms include aqueous solutions, emulsions,suspensions, solutions and/or suspensions reconstituted fromnon-effervescent granules and effervescent preparations reconstitutedfrom effervescent granules. Aqueous solutions include, for example,elixirs and syrups. Emulsions are either oil-in-water or water-in-oil.

Elixirs are clear, sweetened, hydroalcoholic preparations.Pharmaceutically acceptable carriers used in elixirs include solvents.Syrups are concentrated aqueous solutions of a sugar, for example,sucrose, and can include a preservative. An emulsion is a two-phasesystem in which one liquid is dispersed in the form of small globulesthroughout another liquid. Pharmaceutically acceptable carriers used inemulsions are non-aqueous liquids, emulsifying agents and preservatives.Suspensions use pharmaceutically acceptable suspending agents andpreservatives. Pharmaceutically acceptable substances used innon-effervescent granules, to be reconstituted into a liquid oral dosageform, include diluents, sweeteners and wetting agents. Pharmaceuticallyacceptable substances used in effervescent granules, to be reconstitutedinto a liquid oral dosage form, include organic acids and a source ofcarbon dioxide. Coloring and flavoring agents are used in all of theabove dosage forms.

Solvents include glycerin, sorbitol, ethyl alcohol and syrup. Examplesof preservatives include glycerin, methyl and propylparaben, benzoicadd, sodium benzoate and alcohol. Examples of non-aqueous liquidsutilized in emulsions include mineral oil and cottonseed oil. Examplesof emulsifying agents include gelatin, gum arabic, gum tragacanth,xanthan gum, propylene glycol alginate, bentonite, and surfactants suchas polyoxyethylene sorbitan monooleate. Suspending agents includexanthan gum, sodium carboxymethylcellulose, pectin, tragacanth, Veegumand acacia. Diluents include lactose and sucrose. Sweetening agentsinclude sucrose, syrups, glycerin and artificial sweetening agents suchas saccharin. Wetting agents include propylene glycol monostearate,sorbitan monooleate, diethylene glycol monolaurate and polyoxyethylenelauryl ether. Organic acids include citric and tartaric acid. Sources ofcarbon dioxide include sodium bicarbonate and sodium carbonate. Coloringagents include any of the approved certified water soluble FD and Cdyes, and mixtures thereof. Flavoring agents include natural flavorsextracted from plants such fruits, and synthetic blends of compoundswhich produce a pleasant taste sensation.

For a solid dosage form, the solution or suspension, e.g., in propylenecarbonate, vegetable oils or triglycerides, can be encapsulated in agelatin capsule. Such solutions, and the preparation and encapsulationthereof, are disclosed in U.S. Pat. Nos. 4,328,245; 4,409,239; and4,410,545. For a liquid dosage form, the solution, e.g., for example, ina polyethylene glycol, can be diluted with a sufficient quantity of apharmaceutically acceptable liquid carrier, e.g., water, to be easilymeasured for administration.

Alternatively, liquid or semi-solid oral formulations can be prepared bydissolving or dispersing the active compound or salt in vegetable oils,glycols, triglycerides, propylene glycol esters (e.g., propylenecarbonate) and other such carriers, and encapsulating these solutions orsuspensions in hard or soft gelatin capsule shells. Other usefulformulations include those set forth in U.S. Pat. Nos. Re 28,819 and4,358,603. Briefly, such formulations include, but are not limited to,those including a compound provided herein, a dialkylated mono- orpoly-alkylene glycol, including, but not limited to,1,2-dimethoxy-methane, diglyme, triglyme, tetraglyme, polyethyleneglycol-350-dimethyl ether, polyethylene glycol-550-dimethyl ether,polyethylene glycol-750-dimethyl ether wherein 350, 550 and 750 refer tothe approximate average molecular weight of the polyethylene glycol, andone or more antioxidants, such as butylated hydroxytoluene (BHT),butylated hydroxyanisole (BHA), propyl gallate, vitamin E, hydroquinone,hydroxycoumarins, ethanolamine, lecithin, cephalin, ascorbic acid, malicacid, sorbitol, phosphoric acid, thiodipropionic acid and its esters,and dithiocarbamates.

Other formulations include, but are not limited to, aqueous alcoholicsolutions including a pharmaceutically acceptable acetal. Alcohols usedin these formulations are any pharmaceutically acceptable water-misciblesolvents having one or more hydroxyl groups, including, but not limitedto, propylene glycol and ethanol. Acetals include, but are not limitedto, di(lower alkyl) acetals of lower alkyl aldehydes such asacetaldehyde diethyl acetal.

In all embodiments, tablets and capsules formulations can be coated asknown by those of skill in the art in order to modify or sustaindissolution of the active ingredient. Thus, for example, they can becoated with a conventional enterically digestible coating, such asphenylsalicylate, waxes and cellulose acetate phthalate.

Exemplary compositions can include fast-dissolving diluents such asmannitol, lactose, sucrose, and/or cyclodextrins. Also included in suchformulations can be high molecular weight excipients such as cellulosesand microcrystalline celluloses (AVICEL®) or polyethylene glycols (PEG);an excipient to aid mucosal adhesion such as hydroxypropyl cellulose(HPC), hydroxypropyl methyl cellulose (HPMC), sodium carboxymethylcellulose (SCMC), and/or maleic anhydride copolymer (e.g., GANTREZ®);and agents to control release such as polyacrylic copolymer (e.g.,CARBOPOL 934®). Lubricants, glidants, flavors, coloring agents andstabilizers also can be added for ease of fabrication and use.

In certain of such embodiments, a pharmaceutical composition for oraladministration is formulated by combining one or more compounds providedherein with one or more pharmaceutically acceptable carriers. Certain ofsuch carriers enable compounds provided herein to be formulated indosage forms, such as tablets, pills, dragees, capsules, liquids, gels,syrups, slurries and suspensions, for oral ingestion by a subject. Incertain embodiments, pharmaceutical compositions for oral use areobtained by mixing one or more compounds provided herein and one or moresolid excipient. Suitable excipients include, but are not limited to,fillers, such as sugars, including lactose, sucrose, mannitol, orsorbitol; cellulose preparations such as, for example, maize starch,wheat starch, rice starch, potato starch, gelatin, gum tragacanth,methyl cellulose, hydroxypropylmethyl cellulose, sodiumcarboxymethylcellulose, and/or polyvinylpyrrolidone (PVP). In certainembodiments, such a mixture is optionally ground and auxiliaries areoptionally added. In certain embodiments, pharmaceutical compositionsare formed to obtain tablets or dragee cores. In certain embodiments,disintegrating agents (e.g., cross linked polyvinyl pyrrolidone, agar,or alginic acid or a salt thereof, such as sodium alginate) are added.

In certain embodiments, dragee cores are provided with coatings. Incertain of such embodiments, concentrated sugar solutions can be used,which can optionally include gum arabic, talc, polyvinyl pyrrolidone,carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquersolutions, and suitable organic solvents or solvent mixtures. Dyestuffsor pigments can be added to tablets or dragee coatings.

In certain embodiments, a daily dosage regimen for a subject includes anoral dose of between 0.1 μg and 2000 mg of a compound provided herein.In certain embodiments, a daily dosage regimen for a subject includes anoral dose of between 1 μg and 500 mg of a compound provided herein. Incertain embodiments, a daily dosage regimen for a subject includes anoral dose of between 10 μg and 100 mg of a compound provided herein. Incertain embodiments, a daily dosage regimen for a subject includes anoral dose selected from among 0.01, 0.05, 0.075, 0.1, 0.25, 0.5, 0.75,1, 2, 2.5, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 50, 75, 100, 125, 150,175, 180, 190, 200, 225, 250, 300, 400, 500, 750, 800, 850, 900, 950 and1000 milligrams of a compound provided herein. In certain embodiments, adaily dosage regimen is administered as a single daily dose. In certainembodiments, a daily dosage regimen is administered as two, three, four,or more than four doses.

2. Injectables, Solutions and Emulsions

In certain embodiments, the pharmaceutical composition is prepared fortransmucosal administration. In certain of such embodiments penetrantsappropriate to the barrier to be permeated are used in the formulation.Such penetrants are generally known in the art.

Parenteral administration, generally characterized by injection, eithersubcutaneously, intramuscularly or intravenously also is contemplatedherein. Injectables can be prepared in conventional forms, either asliquid solutions or suspensions, solid forms suitable for solution orsuspension in liquid prior to injection, or as emulsions. Suitableexcipients are, for example, water, saline, dextrose, glycerol,mannitol, 1,3-butanediol, Ringer's solution, an isotonic sodium chloridesolution or ethanol. In addition, if desired, the pharmaceuticalcompositions to be administered also can include minor amounts ofnon-toxic auxiliary substances such as wetting or emulsifying agents, pHbuffering agents, stabilizers, solubility enhancers, and other suchagents, such as for example, mono- or diglycerides, fatty acids, such asoleic acid, sodium acetate, sorbitan monolaurate, triethanolamine oleateand cyclodextrins. Implantation of a slow-release or sustained-releasesystem, such that a constant level of dosage is maintained (see, e.g.,U.S. Pat. No. 3,710,795) also is contemplated herein. Briefly, acompound provided herein is dispersed in a solid inner matrix, e.g.,polymethyl-methacrylate, polybutylmethacrylate, plasticized orunplasticized polyvinylchloride, plasticized nylon, plasticizedpolyethylene-terephthalate, natural rubber, polyisoprene,polyisobutylene, polybutadiene, polyethylene, ethylene-vinylacetatecopolymers, silicone rubbers, polydimethyl-siloxanes, silicone carbonatecopolymers, hydrophilic polymers such as hydrogels of esters of acrylicand methacrylic acid, collagen, cross-linked polyvinylalcohol andcross-linked partially hydrolyzed polyvinyl acetate, that is surroundedby an outer polymeric membrane, e.g., polyethylene, polypropylene,ethylene/propylene copolymers, ethylene/ethyl acrylate copolymers,ethylene/vinylacetate copolymers, silicone rubbers, polydimethylsiloxanes, neoprene rubber, chlorinated polyethylene, polyvinylchloride,vinylchloride copolymers with vinyl acetate, vinylidene chloride,ethylene and propylene, ionomer polyethylene terephthalate, butyl rubberepichlorohydrin rubbers, ethylene/vinyl alcohol copolymer,ethylene/vinyl acetate/vinyl alcohol terpolymer, andethylene/vinyloxy-ethanol copolymer, that is insoluble in body fluids.The compound diffuses through the outer polymeric membrane in a releaserate controlling step. The percentage of active compound included insuch parenteral compositions is highly dependent on the specific naturethereof, as well as the activity of the compound and the needs of thesubject.

Parenteral administration of the compositions includes intravenous,subcutaneous and intramuscular administrations. Preparations forparenteral administration include sterile solutions ready for injection,sterile dry soluble products, such as lyophilized powders, ready to becombined with a solvent just prior to use, including hypodermic tablets,sterile suspensions ready for injection, sterile dry insoluble productsready to be combined with a vehicle just prior to use and sterileemulsions. The solutions can be either aqueous or nonaqueous.

If administered intravenously, suitable carriers include physiologicalsaline or phosphate buffered saline (PBS), and solutions includingthickening and solubilizing agents, such as glucose, polyethyleneglycol, and polypropylene glycol and mixtures thereof.

Pharmaceutically acceptable carriers used in parenteral preparationsinclude aqueous vehicles, nonaqueous vehicles, antimicrobial agents,isotonic agents, buffers, antioxidants, local anesthetics, suspendingand dispersing agents, emulsifying agents, sequestering or chelatingagents and other pharmaceutically acceptable substances.

Examples of aqueous vehicles include Sodium Chloride Injection, RingersInjection, Isotonic Dextrose Injection, Sterile Water Injection,Dextrose and Lactated Ringers Injection. Nonaqueous parenteral vehiclesinclude fixed oils of vegetable origin, cottonseed oil, corn oil, sesameoil and peanut oil. Antimicrobial agents in bacteriostatic orfungistatic concentrations must be added to parenteral preparationspackaged in multiple-dose containers which include phenols or cresols,mercurials, benzyl alcohol, chlorobutanol, methyl and propylp-hydroxybenzoic acid esters, thimerosal, benzalkonium chloride andbenzethonium chloride. Isotonic agents include sodium chloride anddextrose. Buffers include phosphate and citrate. Anti-oxidants includesodium bisulfate. Local anesthetics include procaine hydrochloride.Suspending and dispersing agents include sodium carboxymethylcelluose,hydroxy-propyl methylcellulose and polyvinylpyrrolidone. Emulsifyingagents include Polysorbate 80 (TWEEN® 80). A sequestering or chelatingagent of metal ions include EDTA. Pharmaceutical carriers also includeethyl alcohol, polyethylene glycol and propylene glycol for watermiscible vehicles and sodium hydroxide, hydrochloric acid, citric acidor lactic acid for pH adjustment.

The concentration of the pharmaceutically active compound is adjusted sothat an injection provides an effective amount to produce the desiredpharmacological effect. The exact dose depends on the age, weight andcondition of the subject or animal as is known in the art.

The unit dosage parenteral preparations are packaged in an ampoule, avial or a syringe with a needle. All preparations for parenteraladministration must be sterile, as is known and practiced in the art.

Illustratively, intravenous or intraarterial infusion of a sterileaqueous solution including an active compound is an effective mode ofadministration. Another embodiment is a sterile aqueous or oily solutionor suspension including an active material injected as necessary toproduce the desired pharmacological effect.

Injectables are designed for local and systemic administration.Typically a therapeutically effective dosage is formulated to include aconcentration of at least about 0.1% w/w up to about 90% w/w or more, insome embodiments more than 1% w/w, of the active compound to the treatedtissue(s). The active ingredient can be administered at once, or can bedivided into a number of smaller doses to be administered at intervalsof time. The precise dosage and duration of treatment is a function ofthe tissue being treated and can be determined empirically using knowntesting protocols or by extrapolation from in vivo or in vitro testdata. It is to be noted that concentrations and dosage values also canvary with the age of the individual treated. It is to be furtherunderstood that for any particular subject, specific dosage regimensshould be adjusted over time according to the individual need and theprofessional judgment of the person administering or supervising theadministration of the formulations, and that the concentration rangesset forth herein are exemplary only and are not intended to limit thescope or practice of formulations provided herein.

The compounds can be formulated in any suitable vehicle or form. Forexample, they can be in micronized or other suitable form and/or can bederivatized to produce a more soluble active product or to produce aprodrug or for other purposes. The form of the resulting mixture dependsupon a number of factors, including, for example, an intended mode ofadministration and the solubility of the compound in the selectedcarrier or vehicle. The effective concentration is sufficient forameliorating the symptoms of the condition and can be empiricallydetermined.

In certain embodiments, a pharmaceutical composition is prepared foradministration by injection wherein the pharmaceutical compositionincludes a carrier and is formulated in aqueous solution, such as wateror physiologically compatible buffers such as Hanks's solution, Ringer'ssolution, or physiological saline buffer. In certain embodiments, otheringredients are included (e.g., ingredients that aid in solubility orserve as preservatives). In certain embodiments, injectable suspensionsare prepared using appropriate liquid carriers and/or suspending agents.Certain pharmaceutical compositions for injection are presented in unitdosage form, e.g., in ampules or in multi dose containers. Certainpharmaceutical compositions for injection are suspensions, solutions oremulsions in oily or aqueous vehicles, and can include formulatoryagents such as suspending, stabilizing and/or dispersing agents. Certainsolvents suitable for use in pharmaceutical compositions for injectioninclude, but are not limited to, lipophilic solvents and fatty oils,such as sesame oil, synthetic fatty acid esters, such as ethyl oleate ortriglycerides, and liposomes. Aqueous injection suspensions can includesubstances that increase the viscosity of the suspension, such as sodiumcarboxymethyl cellulose, sorbitol, or dextran. Optionally, suchsuspensions also can include suitable stabilizers or agents thatincrease the solubility of the compounds to allow for the preparation ofhighly concentrated solutions.

In certain embodiments, the pharmaceutical composition is prepared foradministration by inhalation. Certain of such pharmaceuticalcompositions for inhalation are prepared in the form of an aerosol sprayin a pressurized pack or a nebulizer. Certain of such pharmaceuticalcompositions include a propellant, e.g., dichlorodifluoromethane,trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide orother suitable gas. In certain embodiments using a pressurized aerosol,the dosage unit can be determined with a valve that delivers a meteredamount. In certain embodiments, capsules and cartridges for use in aninhaler or insufflator can be formulated. Certain of such formulationsinclude a powder mixture of a compound provided herein and a suitablepowder base such as lactose or starch.

In certain embodiments, the pharmaceutical compositions provided areadministered by continuous intravenous infusion. In certain of suchembodiments, from 0.01 μg to 500 mg of the composition is administeredper day.

3. Lyophilized Powders

Of interest herein also are lyophilized powders, which can bereconstituted for administration as solutions, emulsions and othermixtures. They also can be reconstituted and formulated as solids orgels.

The sterile, lyophilized powder is prepared by dissolving a compoundprovided herein, or a pharmaceutically acceptable derivative thereof, ina suitable solvent. The solvent can include an excipient which improvesthe stability or other pharmacological component of the powder orreconstituted solution, prepared from the powder. Excipients that can beused include, but are not limited to, dextrose, sorbitol, fructose, cornsyrup, xylitol, glycerin, glucose, sucrose or other suitable agent. Thesolvent also can include a buffer, such as citrate, sodium or potassiumphosphate or other such buffer known to those of skill in the art at,typically, about neutral pH. Subsequent sterile filtration of thesolution followed by lyophilization under standard conditions known tothose of skill in the art provides the desired formulation. Generally,the resulting solution will be apportioned into vials forlyophilization. In some embodiments, each vial includes a single dosageof from 10 μg to 1000 mg. In another embodiment, each vial includes asingle dosage of from 100 μg to 500 mg. In another embodiment, each vialincludes a single dosage of from 0.1 mg to 50 mg. In another embodiment,each vial includes a single dosage of from 0.5 mg to 20 mg. In anotherembodiment, each vial includes a single dosage of 1 mg, 2 mg, 3 mg, 4mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg or 10 mg. In another embodiment, eachvial includes multiple dosages of the compound. The lyophilized powdercan be stored under appropriate conditions, such as at about 4° C. toroom temperature.

Reconstitution of this lyophilized powder with water for injectionprovides a formulation for use in parenteral administration. Forreconstitution, about 1 mg to 50 mg is added per mL of sterile water orother suitable carrier. In some embodiments, 5 mg to 35 mg is added permL of sterile water or other suitable carrier. In other embodiments, 10mg to 30 mg of lyophilized powder is added per mL of sterile water orother suitable carrier. The precise amount depends upon the selectedcompound. Such amount can be empirically determined.

4. Topical Administration

Topical mixtures are prepared as described for the local and systemicadministration. The resulting mixture can be a solution, suspension,emulsions or the like and are formulated as creams, gels, ointments,emulsions, solutions, elixirs, lotions, suspensions, tinctures, pastes,foams, aerosols, irrigations, sprays, suppositories, bandages, dermalpatches or any other formulations suitable for topical administration.Transdermal skin patches useful for administering the compoundsdisclosed herein include those well known to those of ordinary skill inthat art.

The compounds or pharmaceutically acceptable derivatives thereof can beformulated as aerosols for topical application, such as by inhalation(see, e.g., U.S. Pat. Nos. 4,044,126, 4,414,209, and 4,364,923, whichdescribe aerosols for delivery of a steroid useful for treatment ofinflammatory diseases, particularly asthma). These formulations foradministration to the respiratory tract can be in the form of an aerosolor solution for a nebulizer, or as a microfine powder for insufflation,alone or in combination with an inert carrier such as lactose. In such acase, the particles of the formulation will typically have diameters ofless than 50 microns, in some embodiments less than 10 microns.

In certain embodiments, the pharmaceutical compositions for inhalationare prepared in the form of an aerosol spray in a pressurized pack or anebulizer. Certain of such pharmaceutical compositions include apropellant, e.g., dichlorodifluoro-methane, trichlorofluoromethane,dichlorotetrafluoroethane, carbon dioxide or other suitable gas. Incertain embodiments using a pressurized aerosol, the dosage unit can bedetermined with a valve that delivers a metered amount. In certainembodiments, capsules and cartridges for use in an inhaler orinsufflator can be formulated. Certain of such formulations include apowder mixture of a compound provided herein and a suitable powder basesuch as lactose or starch.

Exemplary compositions for nasal aerosol or inhalation administrationinclude solutions that can include, for example, benzyl alcohol or othersuitable preservatives, absorption promoters to enhance absorptionand/or bioavailability, and/or other solubilizing or dispersing agentssuch as those known in the art.

The compounds can be formulated for local or topical application, suchas for topical application to the skin and mucous membranes, such as inthe eye, in the form of gels, creams, and lotions and for application tothe eye or for intracisternal or intraspinal application. Topicaladministration is contemplated for transdermal delivery and also foradministration to the eyes or mucosa, or for inhalation therapies. Nasalsolutions of the active compound alone or in combination with otherpharmaceutically acceptable excipients also can be administered. Thesesolutions, particularly those intended for ophthalmic use, can beformulated as 0.01%-10% isotonic solutions, pH about 5-7, withappropriate salts. In certain embodiments in which the compositions isadministered locally, the dosage regimen is adjusted to achieve adesired local concentration of a compound provided herein.

In certain embodiments, the pharmaceutical composition is prepared fortopical administration. Certain of such pharmaceutical compositionsinclude bland moisturizing bases, such as ointments or creams. Any ofthe ointment bases known in the art, including water in oil emulsionbases, oil in water emulsion bases, absorption bases, oleaginous basesand water soluble or water miscible bases can be used (e.g., seeRemington: The Science and Practice of Pharmacy, 19th Ed. (Easton, Pa.:Mack Publishing Co., 1995) at pages 1399-1404). Oleaginous ointmentbases are generally anhydrous and include, for example, vegetable oils,animal fats, and semisolid petroleum-based hydrocarbons. Emulsifiableointment bases, also known as absorbent ointment bases, contain littleor no water and include, for example, hydroxystearin sulfate, anhydrouslanolin and hydrophilic petrolatum. Emulsion ointment bases are eitherwater-in-oil (W/O) emulsions or oil-in-water (O/W) emulsions, andinclude, for example, cetyl alcohol, glyceryl monostearate, lanolin,stearic acid and polyethylene glycols of varying molecular weight.Creams are viscous liquids or semi-solid emulsions, and can be eitheroil-in-water or water-in-oil emulsions. Cream bases are water-washable,and contain an oil phase, an emulsifier, and an aqueous phase, which caninclude a fatty alcohol. The emulsifier in a cream formulation isgenerally a nonionic, anionic, cationic, or amphoteric surfactant.Lotions are preparations to be applied to the skin surface withoutfriction, and often include a water or alcohol base, and include anemulsion and often solid particles (such as cocoa butter or fatty acidalcohols).

Exemplary suitable ointment bases include, but are not limited to,petrolatum, petrolatum plus volatile silicones, and lanolin. Creambases, such as those including an emulsion of water, a mineral oil orpetrolatum, one or more fatty alcohols or fatty esters, apolyoxyethylene ether or ester surfactant or polysorbate surfactant,also can be used. Exemplary suitable cream bases include, but are notlimited to, cold cream (USP), hydrous lanolin and hydrophilic ointment(USP). The moisturizing bases can further contain various otheremollients, emulsifiers, perfumes, colorants and preservatives.

Suitable water-in-oil emulsions are commercially available, e.g., blendsof petrolatum, mineral oil, ceresin, lanolin alcohol, panthenol,glycerin and bisabolol under the designation Aquaphor™, available fromBeiersdorf Futuro Inc. (Cincinnati, Ohio); blends of water, glycerin,panthenol, caprylic/capric triglyceride, dicaprylyl carbonate,octyl-dodecanol, C12-15 alkyl benzoate, dimethicone, squalane, tapiocastarch, cetearyl alcohol, glyceryl stearate citrate, myristyl myristate,butylene glycol, benzyl alcohol, carbomer, phenoxyethanol, ammoniumacryloyldimethyltaurate/VP copolymer, sodium hydroxide, methylparaben,propylparaben, iodopropynl butylcarbamate, such as Eucerin™, availablefrom Beiersdorf Futuro Inc. (Cincinnati, Ohio), blends of water, mineraloil, petrolatum, glycerin, isohexadecane, microcrystalline wax, lanolinalcohol, paraffin, panthenol, magnesium sulfate, decyl oleate,octyldodecanol, aluminum stearate, methylchloroisothiazolinone,methylisothiazolinone, citric acid and magnesium stearate, such asNivea™ Cream, available from Beiersdorf Futuro Inc. (Cincinnati, Ohio).

Suitable oil-in-water emulsions are commercially available, e.g., water,mineral oil, petrolatum; sorbitol, stearic acid, lanolin, lanolinalcohol, cetyl alcohol, glyceryl stearate/PEG-100 stearate,triethanolamine, dimethicone, propylene glycol, microcrystalline wax,tri(PPG-3 myristyl ether) citrate, disodium EDTA, methylparaben,ethylparaben, propylparaben, xanthan gum, butylparaben and methyldibromoglutaronitrile, such as Lubriderm™ Cream, available from Pfizer (MorrisPlains, N.J.); a blend of purified water, petrolatum, mineral oil,cetostearyl alcohol, propylene glycol, sodium laurel sulfate, isopropylpalmitate, imidazolidinyl urea, methylparaben and propylparaben, such asDermabase™ cream, available from Paddock Industries, Inc. (Minneapolis,Minn.); and a blend of purified water, glycerin, hydrogenatedpolyisobutene, cetearyl alcohol and ceteareth-20, macadamia nut oil,dimethicone, tocopheryl acetate, stearoxytrimethylsilane (and) stearylalcohol, panthenol, farnesol, benzyl alcohol, phenoxyethanol,acrylates/C10-30 alkyl acrylate crosspolymer, sodium Hydroxide andcitric acid, such as Cetaphil™ lotion, available from GaldermaLaboratories (Ft. Worth, Tex.).

In certain embodiments, the formulation, route of administration anddosage for the pharmaceutical composition provided herein can be chosenin view of a particular subject's condition (see e.g., Fingl et al.,“The Pharmacological Basis of Therapeutics”, Chapter 1, p. 1 (1975)). Incertain embodiments, the pharmaceutical composition is administered as asingle dose. In certain embodiments, a pharmaceutical composition isadministered as a series of two or more doses administered over one ormore days.

5. Compositions for Other Routes of Administration

In certain embodiments, the pharmaceutical composition is prepared fortopical administration such as rectal administration. The pharmaceuticaldosage forms for rectal administration include, but are not limited torectal suppositories, capsules and tablets for systemic effect. Incertain embodiments, a pharmaceutical agent is prepared for rectaladministration, such as a suppositories or retention enema. Certain ofsuch pharmaceutical agents include known ingredients, such as cocoabutter and/or other glycerides. Rectal suppositories are used hereinmean solid bodies for insertion into the rectum which melt or soften atbody temperature releasing one or more pharmacologically ortherapeutically active ingredients. Pharmaceutically acceptablesubstances utilized in rectal suppositories are bases or vehicles andagents to raise the melting point. Examples of bases include cocoabutter (theobroma oil), glycerin-gelatin, Carbowax (polyoxyethyleneglycol) and appropriate mixtures of mono-, di- and triglycerides offatty acids. Combinations of the various bases can be used. In certainembodiments, the pharmaceutical compositions include moisturizing bases,such as ointments or creams. Agents to raise the melting point ofsuppositories include spermaceti and wax. Rectal suppositories can beprepared either by the compressed method or by molding. The typicalweight of a rectal suppository is about 2 to 3 gm.

Tablets and capsules for rectal administration are manufactured usingthe same pharmaceutically acceptable substances and by the same methodsas for formulations for oral administration.

F. ARTICLES OF MANUFACTURE

The compounds provided herein or pharmaceutically acceptable derivativesor prodrugs, or pharmaceutical compositions that include such compounds,can be packaged as articles of manufacture including packaging material,within the packaging material a compound or pharmaceutically acceptablederivative thereof provided herein, which is effective for modulatingthe activity of androgen receptor, or for treatment, prevention oramelioration of one or more symptoms of androgen receptor mediateddiseases or disorders, or diseases or disorders in which androgenreceptor activity is implicated, and a label that indicates that thecompound or composition is used for modulating the activity of androgenreceptor or for treatment, prevention or amelioration of one or moresymptoms of androgen receptor mediated diseases or disorders, ordiseases or disorders in which androgen receptor activity is implicated.

The articles of manufacture provided herein include packaging materials.Packaging materials for use in packaging pharmaceutical products arewell known to those of skill in the art. See, e.g., U.S. Pat. Nos.5,323,907, 5,052,558 and 5,033,252. Examples of pharmaceutical packagingmaterials include, but are not limited to, blister packs, bottles,tubes, inhalers, pumps, bags, vials, containers, syringes and anypackaging material suitable for a selected formulation and intended modeof administration and treatment. A wide array of formulations of thecompounds and compositions provided herein are contemplated as are avariety of treatments for any disease or disorder in which androgenreceptor activity is implicated as a mediator or contributor to thesymptoms or cause.

In certain embodiments, the pharmaceutical compositions can be presentedin a pack or dispenser device which can include one or more unit dosageforms including a compound provided herein. The pack can for exampleinclude metal or plastic foil, such as a blister pack. The pack ordispenser device can be accompanied by instructions for administration.The pack or dispenser also can be accompanied with a notice associatedwith the container in form prescribed by a governmental agencyregulating the manufacture, use, or sale of pharmaceuticals, whichnotice is reflective of approval by the agency of the form of the drugfor human or veterinary administration. Such notice, for example, can bethe labeling approved by the U.S. Food and Drug Administration forprescription drugs, or the approved product insert. Compositionsincluding a compound provided herein formulated in a compatiblepharmaceutical carrier also can be prepared, placed in an appropriatecontainer, and labeled for treatment of an indicated condition.

G. KITS

The compounds provided herein and compositions that include the providedcompounds also can be provided as kits. Kits can include apharmaceutical composition described herein and an item foradministration. For example, a selective androgen receptor modulatorprovided herein can be supplied with a device for administration, suchas a syringe, an inhaler, a dosage cup, a dropper, or an applicator. Thekit can, optionally, include instructions for application includingdosages, dosing regimens and instructions for modes of administration.Kits also can include a pharmaceutical composition described herein andan item for diagnosis. For example, such kits can include an item formeasuring the concentration, amount or activity of androgen receptor ofa subject.

H. EVALUATION OF THE ACTIVITY OF THE COMPOUNDS

Standard physiological, pharmacological and biochemical procedures areavailable for testing the compounds provided herein to identify thosethat possess activity as selective androgen receptor modulators. Invitro and in vivo assays known in the art can be used to evaluate theactivity of the compounds provided herein as selective androgen receptormodulators. Exemplary assays include, but are not limited to,fluorescence polarization assay, luciferase assay and co-transfectionassay. For example, SARMs can be identified using a series of in vitrocell-assays that profiles ligand mediated activation of AR (e.g., seeU.S. Pat. No. 7,196,076). AR agonistic activity can be determined bymonitoring the ability of the SARM compounds to maintain and/orstimulate the growth of AR-containing tissue such as prostate andseminal vesicles, as measured by weight. AR antagonistic activity can bedetermined by monitoring the ability of the SARM compounds to inhibitthe growth of AR-containing tissue.

The agonist and antagonist effects of SARMs can be measured in non-tumortissues via a series of in vivo rat models in which surrogate endpointsare measured in tissues including, but not limited to, the prostate,seminal vesicle, and levitor ani muscle, as well as the hypothalamicaxis via measurement of plasma luteinizing hormone (LH) levels. TheSprague-Dawley rat model has been used extensively as a model foridentifying selective androgen receptor modulators (SARMs) with in vivopharmacological activity, thereby identifying active, non-steroidalselective androgen receptor modulators that can be useful therapeutics,such as for enhancing muscle, bone, and sexual function and treatingprostate cancer (e.g., see Yin et al., J Pharmacol Exp Ther 304(3):1334-1340 (2003), Jasuja et al., Endocrinology 146(10): 4472-4478(2005), Miner et al., Endocrinology 148(1): 363-373 (2007), Morrissey etal., Journal of Andrology 23(3): 341-351 (2002), Adesanya et al.,Scientific Research and Essay 2(8): 309-314 (2007).

The agonist or antagonist activity of a potential SARM also can bemeasured in a normal, non-tumor cell line. Examples of normal, non-tumorcells lines include, but are not limited to, primary rat prostateepithelial and stromal cells, murine muscle cell line C2C12, primaryguinea pig smooth muscle cells, primary smooth-muscle cells fromimmature (I-PSMC) or adult (A-PSMC) rat penis, primary rabbit smoothmuscle cell line, prostatic smooth muscle cell line PS-1, prostaticsmooth muscle cell line PSMC1, mouse bone cell cultures and osteoblastscells and primary rat seminal vesicle lines SVC-1 and SCV-2 (e.g., seeChen et al., FEBS Letters 491: 91-93 (2001), Gerdes et al.,Endocrinology 139: 3569-3577 (1998), Gonzalez-Cadavid et al., Mol. Cell.Endocrinol. 90: 219-229 (1993), Nemeth et al., J. Andrology 19: 718-724(1998), Ricciardelli I., J. Endocrinol. 140: 373-383 (1994),Sadeghi-Nejad et al., Int. J. Impotence Res. 10: 165-169 (1998), Sarahet al., J. Cell. Physiol. 185: 416-424 (2000), and Tajana et al., EMBOJ. 3: 637-644 (1984) Zhang et al., Prostate 30: 117-129 (1997) andZhuang et al., J. Steroid Biochem. Mol. Biol. 41: 693-696 (1992):

Several surrogate endpoint in vivo assays also can be used to examinethe effects of a potential SARM on the AR pathway. These assays measurethe effects of a potential SARM on normal androgen dependent tissues andfunctions, such as, but not limited to, prostate, seminal vesicle,levator ani muscle, bone, libido, fertility and hypothalamus(measurement of blood LH levels). These assays are widely recognized ashaving a direct correlation to the effects of a potential SARM on the ARpathways in humans. Exemplary surrogate endpoint in vivo assays aredescribed in Ashby et al., J. Appl. Tox. 20: 35-47 (2000), Chen et al.,JPET #75424 (2004), Yamada et al., Tox. Sciences 53: 289-296 (2000),Hamann et al., J. Med. Chem. 41: 623-639 (1998), Furr et al., Eur. Urol29: 83-95 (1996), Broulik et al., Bone 20: 473-475 (1997), Maucher etal., J. Cancer Res. Clin. Oncol. 119: 669-674 (1993), Higuchi et al.,Bioorganic & Medicinal Chemistry Letters 17(19): 5442-5446 (2007); Arjanvan Oeveren et al., Bioorganic & Medicinal Chemistry Letters 17(6):1527-1531 (2007); and Vanderschueren et al., Endocrine Reviews 25(3):389-425 (2004).

Animal models bearing a hormone-dependent tumor also can be used toassess the antagonist activity of a potential SARM against the tumor andthe agonist or antagonist activity against AR-containing normalnon-tumor tissues in the animal. For example, the above surrogateendpoint in vivo assays can be run using a rat bearing anandrogen-dependent rat prostate tumor, such as Dunning R-3327. In thismanner, effects of a SARM on a rat androgen-dependent prostate tumor canbe determined while simultaneously examining the effects of the SARMagent on AR-containing normal non-tumor tissues such as, but not limitedto, prostate, seminal vesicle, and levitor ani muscle as well as effectson the hypothalamic axis via measurements of plasma LH levels. In asimilar fashion, immune compromised nude rats bearing humanandrogen-dependent prostate tumors can be used. In this manner, effectsof a SARM on a human androgen-dependent prostate tumor can be determinedwhile simultaneously examining the effects of the SARM agent on normaltissues such as, but not limited to, prostate, seminal vesicle, andlevitor ani muscle as well as effects on the hypothalamic axis viameasurements of plasma LH levels. In addition, in vivo rat assays can beused to determine the effect of SARMs on libido and reproduction.

In certain embodiments, the compounds provided herein are capable ofmodulating activity of androgen receptor in a “co-transfection” assay(also called a “cis-trans” assay), which is known in the art (see e.g.,Evans et al., Science 240: 889-95 (1988); U.S. Pat. Nos. 4,981,784 and5,071,773; and Pathirana et al., “Nonsteroidal Human ProgesteroneReceptor Modulators from the Marie Alga Cymopolia Barbata,” Mol. Pharm.47: 630-35 (1995)). Modulating activity in a co-transfection assay hasbeen shown to correlate with in vivo modulating activity. Thus, incertain embodiments, such assays are predictive of in vivo activity(see, e.g., Berger et al., J. Steroid Biochem. Molec. Biol. 41: 773(1992)). The in vitro biology of the compounds provided herein can bedetermined using any assay known in the art. For example, theco-transfection assay, as described herein or in the art, can be used.The co-transfection assay provides functional activity expressed asagonist EC₅₀ and antagonist IC₅₀ values.

In certain co-transfection assays, two different co-transfectionplasmids are prepared. In the first co-transfection plasmid, cloned cDNAencoding an intracellular receptor (e.g., androgen receptor) isoperatively linked to a constitutive promoter (e.g., the SV 40promoter). In the second co-transfection plasmid, cDNA encoding areporter protein, such as firefly luciferase (LUC), is operativelylinked to a promoter that is activated by a receptor-dependantactivation factor. Both co-transfection plasmids are co-transfected intothe same cells. Expression of the first co-transfection plasmid resultsin production of the intracellular receptor protein. Activation of thatintracellular receptor protein (e.g., by binding of an agonist) resultsin production of a receptor-dependant activation factor for the promoterof the second co-transfection plasmid. That receptor-dependantactivation factor in turn results in expression of the reporter proteinencoded on the second co-transfection plasmid. Thus, reporter proteinexpression is linked to activation of the receptor. Typically, thatreporter activity can be conveniently measured (e.g., as increasedluciferase production).

Certain co-transfection assays can be used to identify agonists, partialagonists, and/or antagonists of intracellular receptors. In certainembodiments, to identify agonists, co-transfected cells are exposed to atest compound. If the test compound is an agonist or partial agonist,reporter activity is expected to increase compared to co-transfectedcells in the absence of the test compound. In certain embodiments, toidentify antagonists, the cells are exposed to a known agonist (e.g.,androgen for the androgen receptor) in the presence and absence of atest compound. If the test compound is an antagonist, reporter activityis expected to decrease relative to that of cells exposed only to theknown agonist.

An exemplary co-transfection assay is the luciferase reporter assay(Evans, Science 240: 889-895 (1988), Berger et al., J Steroid BiochemMol Biol 41: 733-738 (1992)) can be used to establish the ability of acompound of formulae I, II or III to activate (agonist activity) orrepress (antagonist activity) the ability of the human androgen receptor(hAR) to induce gene expression. A human cell line containingendogenously expressed hAR (e.g., MDA-MB-453 cells, derived from a humanmammary carcinoma, American Tissue Type Culture Collection [ATCC] HTB131) can be transfected with an androgen responsive luciferase reporterplasmid to measure the level of cross-reactivity of the compound onother nuclear receptors (such as the glucocorticoid receptor (GR),estrogen receptor (ER), mineralocorticoid receptor (MR), progesteronereceptor (PR), retinoid receptor (RAR), retinoid X receptor (RXR),peroxisome proliferator activating receptors alpha (PPARα), gamma(PPARγ) and delta (PPARδ), liver X receptor (LXR), farnesyl X receptor(FXR) and the pregnane X receptor (PXR)). In the assay, luciferasereporter plasmids containing the cDNA for firefly luciferase (LUC) underthe control of a conditional promoter containing hormone responseelements recognized by the appropriate nuclear receptor areco-transfected into cells with the nuclear receptor expression plasmid.Any appropriate nuclear receptor expression plasmid known in the art canbe used. Exemplary of such plasmids are the reporter plasmid MMTV-LUC,which contains the mouse mammary tumor virus (MTV) long terminal repeat(LTR), which is a conditional promoter containing hormone responseelements recognized by AR, GR, MR and PR (Giguere et al., Cell 46:645-652 (1986)). The reporter plasmid MTV-ERE5-LUC contains the mouseMTV LTR in which the hormone response elements have been deleted andreplaced with five copies of a 33-base pair estrogen response element(ERE) recognized by ER (McDonnell et al., J Biol Chem 269: 11945-11949(1994)). The reporter plasmid MTV-TREp-LUC contains two copies of athyroid hormone response element (TRE) sequence, recognized by RAR(Bissommette et al., Mol Cell Biol 15(10): 5576-5585 (1995), Umesono etal., Nature 336(6196): 262-265 (1988)). The reporter plasmidCRBP-(2)-tk-LUC contains two copies of a response element sequence fromthe cellular retinoid binding protein (CRBP) promoter, recognized byRXR, linked to the tk promoter (Mangelsdorf et al., Cell 66(3): 555-561(1991)). The promoter plasmid pPREA3-tk-LUC contains a response elementsequence from the acyl CoA oxidase gene, recognized by PPARγ, linked tothe tk promoter (Kliewer et al., Nature 355(6359): 446-449 (1992)). Thereporter plasmid LXRE-tk-LUC contains a response element sequencerecognized by LXR, linked to the tk promoter (Willy et al., Genes Dev9(9): 1033-1045 (1995)). The reporter plasmid EcRE7-tk-LUC contains aresponse element sequence recognized by FXR, linked to the tk promoter(Forman et al., Cell 81(5): 687-693 (1995)). The reporter plasmidCYP3A1-tk-LUC contains a response element from the CYP3A1 promoter thatis recognized by PRX, linked to the tk promoter.

Any receptor plasmid expression plasmid known in the art can be used inthe assay. Exemplary receptor expression plasmids include pRShGR(Giguere et al., Cell 46: 645-652 (1986)), pRShMR (Arriza et al.,Science 237: 268-275 (1987)), pSVhPR-B (Vegeto et al., Cell 69: 703-713(1992)), pRShRXRα (Boehm et al., J Med Chem 37(18): 2930-2941 (1994)),pCMVhPPARγ (Willy et al., Genes Dev 9(9): 1033-1045 (1995)),pCMVGAL4hPPARα, pCMVGAL4hPPARδ, pCMV-LXRα and pCMV-FXRα (Cesario et al.,Mol Endocrinol 15(8): 1360-1369 (2001)). The cotransfections areperformed using known methods (Berger et al., J Steroid Biochem Mol Biol41: 733-738 (1992)). The appropriate recombinant DNA receptor expressionplasmid and the luciferase reporter plasmid for the GR, MR, PR, PAP,RXR, PPARα, PPARγ, PPARδ, LXRα, FXRα and PXRα assays are transientlytransfected into cells using a non-liposomal formulation (e.g., theFuGENE 6 transfection reagent, Roche, Indianapolis, Ind., according tomanufacturer's specifications).

Tissue selective androgen receptor agonists provided herein typicallyhave EC₅₀ values of 1 micromolar or less and efficacy values greaterthan about 50% in a standard AR assay, such as the co-transfection assaydescribed herein. In some embodiments, the EC₅₀ is in the range of 1 to1000 μM. In some embodiments, the EC₅₀ is in the range of 1 to 500 μM.In some embodiments, the EC₅₀ is in the range of 1 to 100 μM. In someembodiments, the EC₅₀ is less than 100 μM. In some embodiments, the EC₅₀is less than 75, 70, 65, 60, 55, 50, 45, 40, 35, 30, 25, 20, 15, 10, 5or 1 μM. In some embodiments, the efficacy values are greater than aboutor at 55%, 50%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 105%, 110%,115%, 120%, 125% or greater. In some embodiments, the compounds providedherein have an EC₅₀ value of 10 μM or less and an efficacy value greaterthan 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100%. In someembodiments, the compounds provided herein have an EC₅₀ value of aboutor 1 μM to about or 5 μM and an efficacy value greater than 50%, 55%,60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100%. In some embodiments, thecompounds provided herein exhibit an EC₅₀ value of no greater than 10micromolar, or no greater than 1 micromolar, or no greater than 100nanomolar, or no greater than 10 nanomolar or no greater than 1nanomolar in an assay for determination of AR receptor antagonistactivity.

Tissue selective androgen receptor antagonists provided herein typicallyhave IC₅₀ values of 1 micromolar or less and efficacy values greaterthan about or 50% in a standard AR assay, such as the co-transfectionassay described herein. In some embodiments, the IC₅₀ is in the range of1 to 1000 μM. In some embodiments, the IC₅₀ is in the range of 1 to 500μM. In some embodiments, the IC₅₀ is in the range of 1 to 100 μM. Insome embodiments, the IC₅₀ is less than 100 μM. In some embodiments, theIC₅₀ is less than 75, 70, 65, 60, 55, 50, 45, 40, 35, 30, 25, 20, 15,10, 5 or 1 μM. In some embodiments, the efficacy values are greater thanabout or at 55%, 50%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 105%,110%, 115%, 120%, 125% or greater. In some embodiments, the compoundsprovided herein have an IC₅₀ value of 10 μM or less and an efficacyvalue greater than 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or100%. In some embodiments, the compounds provided herein have an IC₅₀value of about or at 1 μM to about or at 5 μM and an efficacy valuegreater about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100%.In some embodiments, the compounds provided herein exhibit an IC₅₀ valueof no greater than 10 micromolar, or no greater than 1 micromolar, or nogreater than 100 nanomolar, or no greater than 10 nanomolar or nogreater than 1 nanomolar in an assay for determination of AR receptorantagonist activity.

The compounds provided herein exert their receptor-modulatory effectswith high selectivity. This means that they do not bind to certain otherreceptors (other than AR receptors) with high affinity, but rather onlybind to, activate, or inhibit the activity of other receptors withaffinity constants of greater than 1 micromolar (μM). In someembodiments, the compounds only bind to, activate, or inhibit theactivity of other receptors with affinity constants of greater than 100μM. In some embodiments, the compounds only bind to, activate, orinhibit the activity of other receptors with affinity constants ofgreater than 1000 μM. Because side effects are often due to undesirablereceptor activation or antagonism, the high selectivity of the compoundsprovided herein can reduce the incidence of side effects caused byundesirable activation or antagonism of other one or more otherreceptor(s). The compounds provided herein activate AR-induced geneexpression. This was demonstrated in the cotransfection assay inMDA-MB-453 cells.

In assays measuring the affinity and functional activation of AR bycompounds provided herein, compounds of formula I activated AR-dependentgene expression. For example, Compound 102 demonstrated consistentlyhigher potency than other tested androgen modulator compounds in bindingto AR protein, in activating AR in a functional assay, in stimulatingexpression of a key gene in muscle cells, and in repressing a keyinflammatory modulator in bone cells. Higher potency indicates that asubstance can be administered at a lower dose to achieve equaltherapeutic effect. Use of a lower dose is beneficial because it reducesthe size of the oral dosage form or the number of capsules or tabletsrequired to administer an effective dose, thereby improving patientconvenience and comfort, and lower dose improves the safety of a drug byreducing the amount of drug undergoing metabolism, which can yieldproducts that produce off-target side effects.

1. Effect on Muscle

Skeletal α-actin is a major component of skeletal muscle. Androgens playa role in increasing muscle mass and strength (Mooradian et al., EndocrRev 8: 1-28 (1987); Bhasin et al., J Clin Endocrinol Metab 82:407-413(1997); and Bhasin et al., N Engl J Med 335: 1-7 (1996)). Androgensup-regulate α-actin mRNA in rat levator ani skeletal muscle and henceAR-mediated modulation of skeletal α-actin promoter activity is a markerfor studying androgen regulation of gene expression in muscle. Theeffect the compounds provided herein on muscle can be tested by anymethod known in the art. Such methods include increases in the weight ofthe levator ani muscle, which are indicative of anabolic activity, andare accepted in the art as a reliable index of anabolic activity (e.g.,see Antonio et al., J Appl Physiol 87: 2016-2019 (1999)) and theexpression of myosin heavy chain (MHC) subtypes in skeletal muscle. MHCis the predominant protein in skeletal muscle and is expressed in atissue-specific and developmentally regulated manner (Adams et al., Am JPhysiol. 276(4 Pt 2): R954-961 (1999)). Expression of MHC subtypes canbe examined using RT-PCR of masseter muscle tissue of female rats by themethod of Wright et al. (J Appl Physiol. 83(4): 1389-96 (1997)). Atsteady state, mRNA expression parallels the pattern of MHC proteinexpression. Because transcription of MI-IC mRNA occurs in advance of MHCprotein translation, and the increased sensitivity of RT-PCR compared towestern blotting, rapid changes in mRNA expression can be detected andused to analyze the subtle dynamic effects on muscle metabolism. Ananabolic effect on masseter muscle is demonstrated increasingtranscription of MHC, such as MHC type as compared to the untreatedcontrol. Other assays are known in the art (e.g., see Miner et al.,Endocrinology 148(1): 363-373 (2007); Eisenberg et al., Journal ofPharmacology and Experimental Therapeutics 99(1): 38-44 (1950); Labrieet al., J Endocrinology 184: 427-433 (2005)).

An exemplary assay for measuring AR-mediated gene expression in mousemuscle cells is the human skeletal α-actin assay. A mouse muscle cellline (C2C12, obtained from American Type Cell Culture (ATCC, Rockville,Md.)) can be used in the luciferase reporter assay. An SK α-actin-LUCreporter plasmid is constructed by inserting the skeletal α-actinpromoter sequence into a luciferase reporter vector (e.g., by cloningthe human skeletal α-actin promoter region from −77 to +202 bp fromhuman liver genomic DNA (Clontech, Palo Alti, Calif.) and inserting thesequence into a pGL3-Basic reporter plasmid (Promega, Madison, Wis.)).The C2C12 cell line can be transiently transfected with a human ARexpression plasmid and a luciferase reporter plasmid containing theskeletal α-actin promoter upstream of the luciferase cDNA. The compoundsto be tested are added to the cells after transfection and after 24hours of incubation, the cells are lysed with a detergent-containingbuffer and assayed for luciferase activity. The EC₅₀ is determined fromthe concentration response curve for the compound, and efficacy iscalculated by comparison with the standard androgen agonist,dihydrotestosterone.

2. Effect on Bone

Anabolic activity of the compounds provided herein on bone can be testedby any method known in the art. Such methods include bone formationrate, which can be assessed by osteocalcin level measurement. Plasmaosteocalcin levels can be determined using any method known in the art(e.g., see Koyama et al., J Immunol Methods 139(1): 17-23 (1991)). Acommercially available rat osteocalcin EIA kit is available fromBiomedical Technologies Inc. (Stoughton, Mass.). Additional assaysinclude bone nodule formation assays (Vanderschueren et al., EndocrineReviews 25(3): 389-425 (2004), Beresford et al., 45(2): 163-178 (2005)and use of rodent models of osteoporosis (e.g., see Gowen et al., J ClinInvest. 105: 1595-604 (2000); Pietschmann et al., Exp Gerontol. 42(11):1099-1108 (2007); and Wang et al, Bone. 29(2): 141-148 (2001)). Boneturnover markers also have been demonstrated to be an effective,validated tool for monitoring bone activity. For example, bone alkalinephosphatase, urinary hydroxyproline, serum alkaline phosphatase,tartrate-resistant acid phosphatase, osteocalcin levels, and urinarycalcium-creatinine ratio, are used as bone turnover markers (e.g., Sitet al., Adv Ther. 24(5): 987-995 (2007); Szulc et al., Osteoporos Int.18(11): 1451-61 (2007); and Hannon et al., Cancer Treat Rev. 32 Suppl 1:7-14 (2006)). C-telopeptide is used as a bone resorption marker (e.g.,see Srivastava et al., Calcified Tissue International 66(6): 435-442(2000)).

An exemplary cotransfection assay for measuring AR-mediated effects onbone measures the modulation of IL-6 promoter activity. IL-6 is animportant bone resorption factor (Jilka et al, Science 257: 88-91(1992); Bellido et al., J Clin Invest 95: 2886-2895 (1995)).Overexpression of IL-6 can cause severe bone loss in vivo. In vitrostudies have indicated that cytokines, such as TNFα and IL-1β stronglystimulate IL-production through the induction of NF_(K)B (Kurokouchi etal., J Bone Mineral Res 13: 1290-1299 (1998); Ng et al., J Biol Chem269: 19021-19027 (1994)). It has been reported that androgens regulateIL-6 mRNA by suppressing NFKB binding to its response element (Keller etal., J Biol Chem 271: 26267-26275 (1996)). Thus, the AR-mediatedmodulation of IL-6 promoter activity provides an excellent marker forstudying androgen action in bone. The assay is a cotransfection assaythat measures the ability of compound to repress gene expression via thehuman androgen receptor from the IL-6 promoter using a luciferasereporter assay in human osteoblast cells.

A human osteoblast cell line (Saos-2, available from American Type CellCulture (ATCC, Rockville, Md.)) can be used in the luciferase reporterassay. An IL-6-LUC reporter plasmid can be constructed by inserting theIL-6 promoter sequence into a luciferase reporter vector (e.g., bycloning the human IL-6 promoter region from −232 to +17 bp from humanliver genomic DNA (Clontech, Palo Alti, Calif.) and inserting thesequence into a pGL2-enhancer reporter plasmid (Promega, Madison,Wis.)). The Saos-2 cell line can be transiently transfected with a humanAR expression plasmid and a luciferase reporter plasmid containing theIL-6 promoter upstream of the luciferase cDNA. The compounds to betested are added to the cells after transfection and after 24 hours ofincubation, the cells are lysed with a detergent-containing buffer andassayed for luciferase activity. The EC₅₀ is determined from theconcentration response curve for the compound, and the inhibitory effecton TNFα/IL-1β induction (% inhibition) can be calculated according tothe equation:

${\%\mspace{14mu}{Inhibition}} = {\frac{\begin{bmatrix}{\left( {{{TNF}\;\alpha\text{/}{IL}\text{-}1\beta\mspace{14mu}{control}} - {basal}} \right) -} \\\left( {{{androgen}\mspace{14mu}{treatment}} - {basal}} \right)\end{bmatrix}}{\left\lbrack \left( {{{TNF}\;\alpha\text{/}{IL}\text{-}1\beta\mspace{14mu}{control}} - {basal}} \right) \right.} \times 100}$where

-   -   TNFα/IL-1β control=mean luciferase response measured in cells        treated with media containing TNFα and IL-1β without test        compound;    -   basal=mean luciferase response measured in cells treated with        media alone; and    -   androgen treatment=mean luciferase response measured in cells        treated with media containing TNFα and IL-1β with test compound.

3. Antagonist Activity Against Hormone-Dependent Tumors

Various methods for identifying SARMs having antagonist activity againsthormone-dependent tumors while exhibiting no activity, or agonistactivity against other non-tumor tissues containing the androgenreceptor, can be used. For example, antagonist activity inhormone-dependent tumors can be ascertained by screening the compoundsin hormone-dependent tumor cell lines for inhibition of growth, eitherin vitro or in vivo. Examples of hormone-dependent tumor cell lines thatcan be used for screening potential SARMs include, but are not limitedto, human breast tumor cell line MDA MB453, human breast tumor cell lineZR-75-1, murine breast line Shionogi, rat prostate adenocarcinoma lineDunning R-3327, human prostate tumor cell line MDA PCa 2a and PCa 2b,human prostate cell line LNCap, human prostate tumor cell line CWR22,human prostate tumor cell line LuCaP 35 and LuCaP 23.12, human prostatecell line LAPC-4 and LAPC-9, human prostate tumor cell line PC-295,human prostate tumor cell line PC-310, and human osteosarcoma cell lineMG-63. These human and murine prostate and breast cell lines and thetumor model systems derived therein are well accepted by those of skillin the art as indicative of the pharmacology of human hormone-dependenttumors, such as prostate cancer. Examples of the relationship of suchmodels to the human disease state is discussed in the art (e.g., seeJacques et al., Endocrinology 140: 416-421 (1999); Yeap et al.Endocrinology 140: 3282-3291 (1999), Sharma et al., Oncogene 18:5349-5355 (1999), Bentei et al., In Vitro Cell Dev. Biol. 35: 655-662(1999), Suzuki et al., J. Steroid Biochem. Mol. Biol. 37: 559-567(1990), Peehl, Urol. Oncol. 2: 100-102 (1996), Wytske et al., Urol.Oncol. 2: 122-125 (1996), Leland, Urol. Oncol. 2: 126-128 (1996), Buhleret al., The Prostate 43: 63-70 (2000), Navone et al., Clin. Cancer Res.6: 1190-1197 (2000), Etreby et al., The Prostate 42: 99-106 (2000),Jongsma et al., Cancer Res. 60: 741-748 (2000), Jongsma et al., Amer. J.Path. 154: 543-551 (1999), Ye et al., Clin. Cancer Res. 5: 2171-2177(1999), Navone et al., Clin. Cancer Res. 3: 2493-2500 (1997), Chen etal., Cancer Res. 58: 2777-2783 (1998), and Craft et al., Cancer Res. 59:5030-5036 (1999).

4. Efficacy and Toxicity

Compounds disclosed herein can be evaluated for efficacy and toxicityusing known methods. For example, the toxicology of a particularcompound, or of a subset of the compounds, sharing certain chemicalmoieties, may be established by determining in vitro toxicity towards acell line, such as a bacterial or mammalian, including human, cell line.The results of such studies are often predictive of toxicity in animals,such as mammals, or more specifically, humans. Exemplary assays includebacterial reverse mutation assays (e.g., see Ames et al., MutationResearch 31: 347-364 (1975); Green et al., Mutation Research 38: 3-32(1976); and Maron et al., Mutation Research 113: 173-215 (1983),cytological methods for detecting chemical mutagens (Evans, ChemicalMutagens, Principles and Methods for their Detection, Vol. 4 (PlenumPress, New York, N.Y. (1976))), chromosomal aberration assays (Gallowayet al., Mutation Research 312(3): 241-261 (1994)), genotoxicity assays(Federal Register 61: 18198-18202 (1996) and Federal Register 62:16026-16030 (1997)) and cytogenetic assays (Preston et al., MutationResearch 87: 143-188 (1981)).

Alternatively, the toxicity of particular compounds in an animal model,such as mice, rats, rabbits, or monkeys, may be determined using knownmethods. The efficacy of a particular compound may be established usingseveral recognized methods, such as in vitro methods, animal models, orhuman clinical trials. Non-limiting examples of appropriate in vivoanimal models include castrated male rats or aged male orchidectomizedrats. When selecting a model to determine efficacy, the skilled artisancan be guided by the state of the art to choose an appropriate model,dose, and route of administration, and regime. Human clinical trialsalso can be used to determine the efficacy of a compound in humans.

5. Receptor Binding Assays

The binding of the compounds provided herein to androgen receptor can beassessed using any method known in the art. For example, a baculovirusexpression plasmid including cDNA encoding the human steroidal hormonereceptor protein (AR, PR, GR, MR, or ER) can be prepared using standardtechniques. See e.g., Allegretto et al., J. Biol. Chem. 268: 26625(1993); Srinivasan and Thompson, Mol. Endo. 4: 209 (1990); and D. R.O'Reilly et al., in “Baculovirus Expression Vectors”, D. R. O'Reilly etal., eds., W. H. Freeman, New York, N.Y., pp. 139-179 (1992). In anexemplary method, the expression plasmid is infected together with wildtype Autographa californica multiple nuclear polyhedrosis virus DNA intoSpodopter frugiperda-21 (Sf-21) cells to generate recombinant virusincluding AR cDNA. See, e.g., O'Reilly et al., “Regulation of expressionof a baculovirus ecdysteroid UDP glucosyltransferase gene” inBaculovirus Expression Vectors, WH Freeman, NY, 139-179 (1992) and therecombinant virus including receptor cDNA collected.

The recombinant virus then can be used to infect cells, such as Sf-21cells. The infected Sf-21 cells are incubated for 48 hours and thencollected by centrifugation, resuspended in lysis buffer (50 mMpotassium phosphate buffer, pH 7.0, 10 mM monothioglycerol, 5 mM DTT, 20mM sodium molybdate, 1 mM PMSF, 1 μg/mL aprotinin, and 10 μg/mLleupeptin) and homogenized. The resulting receptor lysates are used inthe binding assays. Binding assay samples can be prepared in separatemini-tubes in a 96-well format using Receptor-Assay Buffer (10%glycerol, 25 mM sodium phosphate, 10 mM potassium fluoride, 10 mM sodiummolybdate, 0.25 mM CHAPS, 2 mM DTT and 1 mM EDTA, (adjusted to pH 7.5))containing 50 μg of receptor lysate; 2-4 nM of [³H]-steroid(dihydrotestosterone, progesterone, dexamethasone, aldosterone, orestradiol) at 50-100 Ci/mmol; and either a reference compound or a testcompound. Test compounds included selective androgen receptor bindingcompounds as provided herein. Reference compounds were unlabeledsteroids, which have been previously shown to bind to the steroidhormone receptors, such as dihydrotestosterone for androgen receptors.Each reference compound and test compound are assayed at varyingconcentrations, e.g., ranging from 3.2×10⁻¹⁰ to 10⁻⁵ M and the assaysamples are incubated, e.g., for 16-24 hours at 4° C.

After incubation, 200 μL of 6.25% hydroxylapatite in assay buffer isadded to each assay sample to precipitate the protein. The assay samplesthen are centrifuged, the resulting pellets washed twice with assaybuffer lacking DTT and the radioactivity in counts per minute (CPM) ofeach washed pellet is determined by liquid scintillation counter(MicroBeta™, Wallach).

Specific binding for a particular sample is calculated using theequation:(Sample CPM)−(Average Non-specific CPM)“Average Non-specific CPM” is the amount of radioactivity from samplesincluding an excess (e.g., 1000 nM) of unlabeled steroid, such asdihydrotestosterone. IC₅₀ values (the concentration of test compoundrequired to decrease specific binding by 50%) were determined using thelog-logit (Hill) method.

In some embodiments, the compounds provided herein exhibit a K_(i) of nogreater than 10 micromolar, or no greater than 5 micromolar, or nogreater than 1 micromolar, or no greater than 100 nanomolar, or nogreater than 10 nanomolar or no greater than 1 nanomolar in an ARreceptor binding assay.

6. In Vivo Assay—Sprague-Dawley Rat Model

The Sprague-Dawley rat model is used in the art as a model foridentifying selective androgen receptor modulators (SARMs) that exhibitin vivo pharmacological activity, thereby identifying active,nonsteroidal selective androgen receptor modulators. In the rat, theeffects of androgens in skeletal muscle can be assessed by monitoringthe weight of the levator ani muscle (Herschberger et al., Proc Soc ExpBiol Med 83: 175-180 (1981)), a muscle that expresses high levels ofandrogen receptor (Max et al., Biochem J 200: 77-82 (1981)). Underandrogen treatment, the weight of the levator ani muscle markedlyincreases, providing a reliable endpoint to study the anabolic effect ofandrogens in skeletal muscle (Herschberger et al., Proc Soc Exp Biol Med83: 175-180 (1981)).

Androgens also stimulate sebaceous gland secretions in the skin and havebeen linked with increased sebum production and acne (Boudou et al., JClin Endocrinol Metab 80(4): 1158-1161 (1995)). The preputial gland inrodents is a modified sebaceous gland and has been used as an indicatorof sebum production (Miyake et al., J Invest Dermatol 103: 721-725(1994); Nickerson et al., Acta Anat (Basel) 94: 481-489 (1976); andDeplewski et al., Endocrinology 138: 4416-4420 (1997)). Androgensupplementation increases the weight of sebaceous glands and theirsecretions.

Male steroid hormones also have been found to be critical formaintenance and growth of bone. For example, osteopenia often occurs inhypogonadism in men, a condition that results in reduced circulatingtestosterone levels (Devogelaer et al., Maturitas 15: 17-23 (1992)).Osteopenia is a degenerative process that can be reversed withtestosterone supplementation (Finkelstein et al., J Clin Endocrinol &Metabolism 69: 776-783 (1989); Behre et al., J Clin Endocrinol &Metabolism 82: 2386-2390 (1997); and Katznelson et al., J ClinEndocrinol & Metabolism 81: 4358-4365 (1996)). A similar phenomenon canbe observed in male rats in which bone loss due to castration can berestored by administering androgen (Wakley et al., J Bone & MineralResearch 6: 325-330 (1991)). In order to assess bone loss in rats,biochemical markers, such as serum osteocalcin, can be measured.Osteocalcin is a bone matrix protein component that is synthesized anddeposited by bone-forming osteoblasts and it is also released from thebone during bone resorption by osteoclasts (Ivaska et al., J Biol Chem279: 18361-18369 (2004)). During both processes, this protein and itsbreakdown products are released into the circulation and can served asbiochemical markers. Measurement of circulating osteocalcin levels inthe serum of rats can by used to assess bone turnover or metabolism(Vanderschueren et al., Endocrinology 141(5): 1642-1647 (2000);Vanderschueren et al., Bone Miner 26(2): 123-131 (1994); and Hunter etal., Arthritis Res Ther 10(4): R102 (2008)).

Mature male Sprague-Dawley rats of approximately 2 months of agegenerally are used. After a one-week acclimation period, they arecastrated under isoflurane anesthesia. One group of animals issham-operated and treated with vehicle and serves as a control. Aftersurgery, the rats are sorted into groups so that no statisticallysignificant differences in body weight are observed. Treatment generallybegins the same day of the surgery after the surgical procedure. Animalsare treated either orally (4 mL/kg) or subcutaneously (0.4 mL/kg) eachmorning for 14 consecutive days with vehicle or various compoundsolutions of different strengths. Approximately 24 hours after the lastdose, rats are sacrificed by decapitation and organs and bloodcollected. Trunk blood is collected and then the ventral prostate,seminal vesicles and levator ani muscle are dissected out, blotted dryand weighed individually. The blood is allowed to clot at roomtemperature and the serum separated by centrifugation. The serum is thencollected and kept frozen until processed for measurement of LH levelsby radioimmunoassay (RIA).

The compound of interest is suspended in a vehicle, such as a solutionof 9.995% polyethylene glycol (average molecular weight 400; PEG-400;Sigma, St. Louis, Mo.), 0.005% Tween 80 (polyoxyethylene sorbitanmonooleate; Sigma, St. Louis, Mo.), and 90.0% of a 1%carboxymethylcellulose (CMC; Sigma, St. Louis, Mo.) solution in NanoPurewater. The compound is suspended in the vehicle and can be homogenizedin a blender for 4 minutes. The suspension also can be sonicated for 2minutes. The high concentration formulation was then diluted usingvehicle to obtain the proper volume and concentrations of the dosingmaterials.

Testosterone is dissolved in a 30:70 mixture of PEG-400 and DMSO andadministered subcutaneously.

Serum samples are assayed for LH with a double antiserum procedure usingreagents from the National Institute of Diabetes and Digestive andKidney Diseases (NIDDK). In brief, samples and standards(NIDDK-rLH-RP-3) in a total volume of 200 μl are incubated at roomtemperature for 2-3 days with 100 μl primary antiserum (rabbitNIDDK-anti-rLH-S-11) diluted 1:100,000. Thereafter, 100 μl of iodinatedLH (Covance Laboratories Inc.) diluted to 200,000-300,000 cpm/ml isadded to the tubes and incubation continued for an additional 24 hourperiod. Bound hormone is separated from free hormone by precipitationwith a specific goat anti-rabbit serum (GARS; Antibodies Inc). For thispurpose, 50 μl of 4% normal rabbit serum is added to each incubationtube, after which an additional 50 μl of a 1:10 GARS solution is added.The tubes are vortexed and incubated overnight at 4° C. The assay isterminated by centrifugation at 2,500 rpm for 30 min in a centrifuge at4° C. The supernatants are decanted and discarded and the pellets arecounted in a 10-channel gamma-counter. The assay has a minimaldetectable amount of 0.001 ng/tube and the intra- and inter-assayvariability is less than 10%. In order to minimize interassayvariability, all samples from a single study are run in the same assay.

Results are analyzed by analysis of variance on Box-Cox transformed data(Box and Cox, J Roy Statist Soc Series B 26: 211-252 (1964), Box andHill, Technometrics 16: 385-389 (1974) and Peltier et al., J. Anim. Sci.76: 847-849 (1998)). For ventral prostate and LH levels the logarithm ofthe data are used for statistical analysis. Seminal vesicle and levatorani muscle data are elevated to 0.2 and 0.6, respectively. Thesetransformations are performed to ensure that variances are homogeneousamong groups and that the residuals of the one-way analysis of variancemodel followed a Gaussian (normal) distribution. When the analyses ofvariance reach significance, data are further evaluated by the Dunnett'stest. A P<0.05 is considered as the minimum criterion to declarestatistically significant differences.

In addition, efficacy data are calculated as percentage of the responseobserved in the sham-operated group. In this respect, theorchidectomized, vehicle-treated group is considered as 0% efficacy,whereas the sham-operated group represented 100% efficacy. Thisconversion allows direct comparison of data independently of havingdifferent controls as is the case in experiments using subcutaneouslyand orally administered compounds.

Potencies are estimated on the transformed data using a four-parameterlogistic equation. The model estimates EC₅₀'s in a logarithmic scale,since log EC₅₀ is a more robust estimate of the potency (Ghosh et al., JBiopharm Stat 8: 645-665 (1998)). The model also provides a SE for theestimate that is used to calculate 95% confidence limits of theestimated potencies. The following equation exemplifies the basic,reparametrized four parameter logistic equation used in estimatingpotencies in these studies:

$\frac{A - D}{1 + e^{B \cdot {({{logC} - {logx}})}}} + D$where A is the maximum, D is the minimum, B is the slope, C is eitherthe EC₅₀ or the IC₅₀ depending on the direction of the response, and xis the dose of the compound used.

I. METHODS OF USE OF THE COMPOUNDS AND COMPOSITIONS

Methods of use of the compounds and compositions provided herein alsoare provided. The methods include in vitro and in vivo uses of thecompounds and compositions for altering androgen receptor activity andfor treatment, prevention, or amelioration of one or more symptoms ofdiseases or disorder that are modulated through androgen receptoractivity, or in which androgen receptor activity is implicated. Incertain embodiments, provided herein are methods of treating a subjectby administering a compound provided herein. In certain embodiments,such subject exhibits symptoms or signs of a androgen receptor mediatedcondition. In certain embodiments, a subject is treated prophylacticallyto reduce or prevent the occurrence of a condition.

The compounds provided herein can be used in the treatment of a varietyof conditions. For example, a compound of Formula I, II or III orpharmaceutically acceptable salts or prodrugs thereof can be used totreat a condition including, but not limited to, maintenance of musclestrength and function (e.g., in the elderly); reversal or prevention offrailty or age-related functional decline (“ARFD”) in the elderly (e.g.,sarcopenia); treatment of catabolic side effects of glucocorticoids;prevention and/or treatment of reduced bone mass, density or growth(e.g., osteoporosis and osteopenia); treatment of chronic fatiguesyndrome (CFS); chronic myalgia; treatment of acute fatigue syndrome andmuscle loss following elective surgery (e.g., post-surgicalrehabilitation); accelerating of wound healing; accelerating bonefracture repair (such as accelerating the recovery of hip fracturepatients); accelerating healing of complicated fractures, e.g.distraction osteogenesis; in joint replacement; prevention ofpost-surgical adhesion formation; acceleration of tooth repair orgrowth; maintenance of sensory function (e.g., hearing, sight, olfactionand taste); treatment of periodontal disease; treatment of wastingsecondary to fractures and wasting in connection with chronicobstructive pulmonary disease (COPD), chronic liver disease, AIDS,weightlessness, cancer cachexia, burn and trauma recovery, chroniccatabolic state (e.g., coma), eating disorders (e.g., anorexia) andchemotherapy; treatment of cardiomyopathy; treatment ofthrombocytopenia; treatment of growth retardation in connection withCrohn's disease; treatment of short bowel syndrome; treatment ofirritable bowel syndrome; treatment of inflammatory bowel disease;treatment of Crohn's disease and ulcerative colitis; treatment ofcomplications associated with transplantation; treatment ofphysiological short stature including growth hormone deficient childrenand short stature associated with chronic illness; treatment of obesityand growth retardation associated with obesity; treatment of anorexia(e.g., associated with cachexia or aging); treatment of hypercortisolismand Cushing's syndrome; Paget's disease; treatment of osteoarthritis;induction of pulsatile growth hormone release; treatment ofosteochondro-dysplasias; treatment of depression, nervousness,irritability and stress; treatment of reduced mental energy and lowself-esteem (e.g., motivation/assertiveness); improvement of cognitivefunction (e.g., the treatment of dementia, including Alzheimer's diseaseand short term memory loss); treatment of catabolism in connection withpulmonary dysfunction and ventilator dependency; treatment of cardiacdysfunction (e.g., associated with valvular disease, myocardialinfarction, cardiac hypertrophy or congestive heart failure); loweringblood pressure; protection against ventricular dysfunction or preventionof reperfusion events; treatment of adults in chronic dialysis; reversalor slowing of the catabolic state of aging; attenuation or reversal ofprotein catabolic responses following trauma (e.g., reversal of thecatabolic state associated with surgery, congestive heart failure,cardiac myopathy, burns, cancer, COPD); reducing cachexia and proteinloss due to chronic illness such as cancer or AIDS; treatment ofhyperinsulinemia including nesidioblastosis; treatment ofimmunosuppressed subjects; treatment of wasting in connection withmultiple sclerosis or other neurodegenerative disorders; promotion ofmyelin repair; maintenance of skin thickness; treatment of metabolichomeostasis and renal homeostasis (e.g., in the frail elderly);stimulation of osteoblasts, bone remodeling and cartilage growth;regulation of food intake; treatment of insulin resistance, includingNIDDM, in mammals (e.g., humans); treatment of insulin resistance in theheart; improvement of sleep quality and correction of the relativehyposomatotropism of senescence due to high increase in REM sleep and adecrease in REM latency; treatment of hypothermia; treatment ofcongestive heart failure; treatment of lipodystrophy (e.g., in subjectstaking HIV or AIDS therapies such as protease inhibitors); treatment ofmuscular atrophy (e.g., due to physical inactivity, bed rest or reducedweight-bearing conditions); treatment of musculoskeletal impairment(e.g., in the elderly); improvement of the overall pulmonary function;treatment of sleep disorders; and the treatment of the catabolic stateof prolonged critical illness; treatment of hirsutism, acne, seborrhea,androgenic alopecia, anemia, hyperpilosity, benign prostate hypertrophy,adenomas and neoplasias of the prostate (e.g., advanced metastaticprostate cancer) and malignant tumor cells containing the androgenreceptor, such as is the case for breast, brain, skin, ovarian, bladder,lymphatic, liver and kidney cancers; cancers of the skin, pancreas,endometrium, lung and colon; osteosarcoma; hypercalcemia of malignancy;metastatic bone disease; treatment of spermatogenesis, endometriosis andpolycystic ovary syndrome; counteracting preeclampsia, eclampsia ofpregnancy and preterm labor; treatment of premenstrual syndrome;treatment of vaginal dryness; age related decreased testosterone levelsin men, male menopause, hypogonadism, male hormone replacement, male andfemale sexual dysfunction (e.g., erectile dysfunction, decreased sexdrive, sexual well-being, decreased libido), male and femalecontraception, hair loss, Reaven's Syndrome and the enhancement of boneand muscle performance/strength.

In certain embodiments, provided are methods for treating a subject byadministering one or more than one compound of Formula I, II or III orpharmaceutically acceptable salts or prodrugs thereof. Exemplaryconditions that can be treated with the selective androgen receptormodulators provided herein include, but are not limited to,hypogonadism, wasting diseases, cancer cachexia, frailty, infertility,osteoporosis, hirsutism, acne, male-pattern baldness, prostatichyperplasia, and cancer, including, but not limited to, varioushormone-dependent cancers, including, without limitation, prostate andbreast cancer. In certain embodiments, a selective androgen receptoragonist or partial agonist is used for male hormone replacement therapy.In certain embodiments, one or more selective androgen receptor agonistsand/or partial agonists are used to stimulate hematopoiesis. In certainembodiments, a selective androgen receptor agonist or partial agonist isused as an anabolic agent. In certain embodiments, a selective androgenreceptor agonist and/or partial agonist is used to improve athleticperformance.

In another embodiment, a compound of Formula I, II or III orpharmaceutically acceptable salts or prodrugs thereof is administered toa subject in order to treat a condition responsive to an AR modulatorcompound. The method includes administering to a subject having acondition responsive to an AR modulator compound a therapeuticallyeffective amount of one or more than one compound provided herein totreat the condition responsive to an AR modulator compound. In someembodiments, the condition is treated by agonizing the androgenreceptor. In some embodiments, the condition is treated by antagonizingthe androgen receptor. In various embodiments, the condition treated isselected from among hypogonadism, lower than normal testosterone plasmalevels, infertility, sexual arousal disorder, disorders of libido,muscle wasting, cachexia, sarcopenia, frailty, bone density loss, mooddisorders (including lack of well being, lack of vigor, anger,irritability, sadness, tiredness, nervousness and depression), impairedcognitive function (including verbal fluency and spatial memory),neurodegenerative disorders, including Alzheimer's disease, mildcognition impairment, Lewis body dementia, and frontal temporaldementia, xerophthalmia, metabolic disorders, including dyslipidemia,atherosclerosis, and non-insulin dependent diabetes (NIDDM),cardiovascular disorders including but not limited to hypertension,coronary artery disease, and myocardial perfusion, obesity, anemia,prostate cancer, and schizophrenia. In other embodiments, a compound ofFormula I, II or III or pharmaceutically acceptable salts or prodrugsthereof can be administered to a subject in order to prevent a conditionin the subject. In various embodiments, the condition prevented includesbone density loss, xerophthalmia, metabolic disorders, includingdyslipidemia, atherosclerosis, and non-insulin dependent diabetes(NIDDM), cardiovascular disorders including hypertension, coronaryartery disease, and myocardial perfusion, obesity and prostate cancer.

In certain embodiments, one or more than one compound of Formula I, IIor III or pharmaceutically acceptable salts or prodrugs thereof is usedto treat acne, male-pattern baldness, wasting diseases, hirsutism,hypogonadism, osteoporosis, infertility, impotence, obesity, and cancer.In certain embodiments, one or more compounds provided herein are usedto stimulate hematopoiesis. In certain embodiments, one or morecompounds provided herein are used for contraception.

In certain embodiments, provided herein are methods for treating asubject having a condition caused by androgen deficiency or a conditionameliorated by androgen replacement. The methods include administeringto the subject a therapeutically effective amount of one or more thanone compound of Formula I, II or III or pharmaceutically acceptablesalts or prodrugs thereof, and thereby treating the condition. Incertain embodiments, the condition is selected from among abdominalobesity, Alzheimer's disease, anemia, an arthritic condition,atherosclerosis, benign prostatic hyperplasia (BPH), cancer cachexia,cognitive decline, depression, metabolic syndrome, a muscular dystrophy,obesity, osteopenia, osteoporosis, a periodontal disease, prostatecancer, sexual dysfunction, sleep apnea, type II diabetes, bonefracture, frailty, wasting, aging skin, hypogonadism, post-menopausalsymptoms in women, female sexual dysfunction, premature ovarian failure,atherosclerosis, hypercholesterolemia, hyperlipidemia, aplastic anemiaand other hematopoietic disorders, pancreatic cancer, renal cancer,arthritis and joint repair.

1. Methods of Treating Muscle Wasting

In certain embodiments, provided herein are methods of treating,preventing, suppressing, inhibiting or reducing the incidence of musclewasting in a subject. The methods include administering to the subjectone or more than one compound of Formula I, II or III orpharmaceutically acceptable salts or prodrugs thereof, in an amounteffective to treat, prevent, suppress, inhibit or reduce muscle wastingin the subject. In some embodiments, the muscle wasting is caused by acondition selected from among andropause, a spinal muscular atrophy, amuscular dystrophy, myasthenia gravis, AIDS cachexia, cardiac cachexia,cancer cachexia, cancer, Chronic Obstructive Pulmonary Disease (COPD),emphysema, diabetes, HIV infection, acquired immunodeficiency syndrome(AIDS), sepsis, tuberculosis, renal failure, heart failure,cardiomyopathy, bed rest, disuse, inactivity, microgravity,malnutrition, sarcopenia and aging. In some embodiments, the one or morethan one compound of Formula I, II or III or pharmaceutically acceptablesalts or prodrugs thereof is orally administered to the subject. In onemethod, the compounds provided herein are used in a method for thetreatment of muscular dystrophy, sarcopenia and frailty. In oneembodiment, the methods include co-administering one or more than onecompound provided herein with one or more agents selected from amonginterleukin-10 (IL-10), interleukin-4 (IL-4), a TNF inhibitor,fluorinated 4-azasteroid derivatives, glial growth factors,acetylcholine receptor inducing activity (ARIA), heregulins, neudifferentiation factor, and neuregulins (e.g., see U.S. Pat. Nos.6,444,642 and 7,037,888).

In one embodiment, provided herein are methods of treating amuscle-wasting condition associated with chronic illness. The methodsinclude administering to the subject one or more than one compound ofFormula I, II or III or pharmaceutically acceptable salts or prodrugsthereof, in an amount effective to treat the muscle-wasting condition.In one embodiment, provided herein are methods for preventing a musclewasting disorder in a subject, which include administering to thesubject one or more than one compound of Formula I, II or III orpharmaceutically acceptable salts or prodrugs thereof, in an amounteffective to preventing a muscle wasting disorder in the subject. In oneembodiment, provided herein are methods for suppressing a muscle wastingdisorder in a subject, which include administering to the subject one ormore than one compound of Formula I, II or III or pharmaceuticallyacceptable salts or prodrugs thereof, in an amount effective to suppressthe muscle wasting disorder in a subject. In one embodiment, providedherein are methods for reducing the incidence of a muscle wastingdisorder in a subject, which include administering to the subject one ormore than one compound of Formula I, II or III or pharmaceuticallyacceptable salts or prodrugs thereof, in an amount effective to suppressthe muscle wasting disorder in a subject.

Methods for identifying a subject in need of treatment for a muscularwasting disease are known in the art. For example, a subject in need oftreatment for a muscular wasting disease will often generate lesselectrical activity during muscle contraction as compared to a healthysubject and this can be detected by electromyography. Alternativemethods for diagnosis include, for example, blood tests and musclebiopsies. Suitably, blood tests can be run to determine the levels ofvarious constituents of muscle and muscle fibers. For example, manymuscular wasting diseases can be diagnosed by conducting a blood test tomeasure the level of creatinine in the blood. Creatinine is a breakdownproduct of creatine, which is an important constituent of muscle. Bloodtests for determining the amount of creatine phosphokinase (CPK), whichis an enzyme found predominantly in the heart, brain, and skeletalmuscle, can be conducted to diagnose a subject in need for treatment ofa muscular wasting disease. Specifically, when the total CPK level issubstantially elevated, it usually indicates injury or stress to one ormore of the heart, brain, and skeletal muscle. Subjects that may beaffected by either Duchenne muscular dystrophy or Becker musculardystrophy can be diagnosed by measuring the level of dystrophin.Typically, in subjects with either Duchenne muscular dystrophy or Beckermuscular dystrophy, the level of dystrophin is deficient; but, in asubject with Duchenne muscular dystrophy, the level is more severelydeficient.

Muscle biopsies also can be used to identify a subject in need oftreatment for a muscular wasting disease. Generally, during a musclebiopsy, a small piece of muscle tissue is removed surgically forlaboratory analysis. The analysis can reveal abnormalities in themuscle, such as inflammation, damage, or infection. The subject also canbe diagnosed for a muscular wasting disease using magnetic resonanceimagining (MRI). During an MRI, cross-sectional images of muscle aregenerated by a magnetic field and radio waves. Similar to the musclebiopsy analysis, the image generated by an MRI can reveal abnormalitiesin the muscle, such as inflammation, damage, or infection.

2. Methods of Improving Muscle Performance, Size and/or Strength

In certain embodiments, provided herein are methods of increasing muscleperformance, muscle size, muscle strength, or any combination thereof ina subject. The methods include administering to the subject one or morethan one compound of Formula I, II or III or pharmaceutically acceptablesalts or prodrugs thereof, in an amount effective to increase muscleperformance, muscle size, and/or muscle strength in the subject.

In some embodiments, provided herein are methods of activating thefunction of the androgen receptor muscle tissue and blocking orinhibiting the function of the androgen receptor in the prostate of amale individual or in the uterus of a female individual. The methodsinclude administering to the subject one or more than one compound ofFormula I, II or III or pharmaceutically acceptable salts or prodrugsthereof, in an amount effective to activate the function of the androgenreceptor in muscle tissue and to block or inhibit the function of theandrogen receptor in the prostate of a male individual or in the uterusof a female individual.

3. Methods of Improving Athletic Performance

In certain embodiments, one or more than one compound of Formula I, IIor III or pharmaceutically acceptable salts or prodrugs thereof is usedto improve athletic performance. The methods include administering oneor more than one compound of Formula I, II or III or pharmaceuticallyacceptable salts or prodrugs thereof in a therapeutically effectiveamount to improve athletic performance. In some embodiments, one or morecompounds provided herein are used, for example, to shorten the timenormally needed to recover from physical exertion or to increase musclestrength. Athletes to whom one or more compounds provided herein can beadministered include, but are not limited to, horses, dogs and humans.In certain embodiments, one or more than one compound of Formula I, IIor III or pharmaceutically acceptable salts or prodrugs thereof isadministered to an athlete engaged in a professional or recreationalcompetition, including, but not limited to weight-lifting,body-building, track and field events, and any of various team sports.

4. Methods of Treating Bone-Related Conditions

In certain embodiments, provided herein are methods of treating,preventing, suppressing, inhibiting or reducing the incidence ofosteoporosis, osteopenia, gluco-corticoid-induced osteoporosis or bonefracture in a subject. The methods include administering to the subjectone or more than one compound of Formula I, II or III orpharmaceutically acceptable salts or prodrugs thereof, in an amounteffective to treat osteoporosis, osteopenia, glucocorticoid-inducedosteoporosis or bone fracture in the subject. In one embodiment, the oneor more than one compound of Formula I, II or III or pharmaceuticallyacceptable salts or prodrugs thereof is co-administered with aneffective amount of at least one other therapeutic agent, such as anestrogen or estrogen derivatives, alone or in combination with progestinor progestin derivatives; a bisphosphonate; an anti-estrogen; aselective estrogen receptor modulators (SERM); an α_(v)β₃ integrinreceptor antagonist; a cathepsin inhibitor; a proton pump inhibitor; aPPARγ inhibitor; calcitonin; and osteoprotegerin. In one embodiment, themethod is for the treatment of osteoporosis. In one embodiment, themethod is for the treatment of osteopenia. In one embodiment, the methodis for the treatment of glucocorticoid-induced osteoporosis. In oneembodiment, the method is for the treatment of bone fracture.

In some embodiments, provided herein are methods of activating thefunction of the androgen receptor in bone tissue and blocking orinhibiting the function of the androgen receptor in the prostate of amale individual or in the uterus of a female individual. The methodsinclude administering to the subject one or more than one compound ofFormula I, II or III or pharmaceutically acceptable salts or prodrugsthereof, in an amount effective to activate the function of the androgenreceptor in bone tissue and to block or inhibit the function of theandrogen receptor in the prostate of a male individual or in the uterusof a female individual.

In certain embodiments, provided herein are methods of increasing thestrength of, or mass of a bone of a subject, or for promoting boneformation in a subject. The methods include administering one or morethan one compound of Formula I, II or III or pharmaceutically acceptablesalts or prodrugs thereof in an amount effective to increase thestrength of, or mass of a bone of a subject, or to promote boneformation in a subject.

In some embodiments, provided herein are methods for preventing abone-related disorder in a subject, which include administering one ormore than one compound of Formula I, II or III or pharmaceuticallyacceptable salts or prodrugs thereof in an amount effective to preventthe bone-related disorder in the subject. In some embodiments, providedherein are methods for suppressing a bone-related disorder in a subject,which include administering one or more than one compound of Formula I,II or III or pharmaceutically acceptable salts or prodrugs thereof in anamount effective to suppress the bone-related disorder in the subject.In some embodiments, provided herein are methods for inhibiting abone-related disorder in a subject, which include administering one ormore than one compound of Formula I, II or III or pharmaceuticallyacceptable salts or prodrugs thereof in an amount effective to inhibitthe bone-related disorder in the subject.

In one embodiment, the bone-related disorder is osteoporosis. In anotherembodiment, the bone-related disorder is osteopenia, In anotherembodiment, the bone-related disorder is increased bone resorption. Inanother embodiment, the bone-related disorder is bone fracture. Inanother embodiment, the bone-related disorder is bone frailty. Inanother embodiment, the bone-related disorder is any combination ofosteoporosis, osteopenia, increased bone resorption, bone fracture andbone frailty.

In one embodiment, the osteoporosis results from androgen deprivation.In another embodiment, the osteoporosis follows androgen deprivation. Inanother embodiment, the osteoporosis is primary osteoporosis. In anotherembodiment, the osteoporosis is secondary osteoporosis. In anotherembodiment, the osteoporosis is postmenopausal osteoporosis. In anotherembodiment, the osteoporosis is juvenile osteoporosis. In anotherembodiment, the osteoporosis is idiopathic osteoporosis. In anotherembodiment, the osteoporosis is senile osteoporosis.

5. Methods of Treating Cancer

In certain embodiments, one or more than one compound of Formula I, IIor III or pharmaceutically acceptable salts or prodrugs thereof is usedfor treating, preventing, suppressing, inhibiting or reducing theincidence of cancer in a subject. Certain exemplary cancers include, butare not limited to, breast cancer, colorectal cancer, gastric carcinoma,glioma, head and neck squamous cell carcinoma, skin cancer, papillaryrenal carcinoma, leukemia, lymphoma, Li-Fraumeni syndrome, malignantpleural mesothelioma, melanoma, multiple myeloma, non-small cell lungcancer, synovial sarcoma, thyroid carcinoma, transitional cell carcinomaof urinary bladder, and prostate cancer, including, but not limited toprostatic hyperplasia. The methods include administering one or morecompounds provided herein in a therapeutically effective amount to treatthe cancer. In one embodiment, administration of the one or more thanone compound provided herein to a subject afflicted with a cancerouscondition alleviates the cancerous condition by killing the cancerouscells. In one embodiment, administration of the one or more than onecompound provided herein to a subject afflicted with a cancerouscondition results in the inhibition of growth and/or metastasis of thecancer.

In some embodiments, one or more than one compound of Formula I, II orIII or pharmaceutically acceptable salts or prodrugs thereof isadministered in combination with one or more other therapeutic agents,such as, but not limited to, anti-proliferative agents, such aspaclitaxel, a paclitaxel derivative, taxanes and vinca alkaloids,anti-tumor agents, such as mitomycin C or doxorubicin, hormones andantagonists, such as adreno-corticosteroids (prednisone), progestins(hydroxyprogesterone caproate, medroprogesterone acetate and megestrolacetate), estrogens (diethylstilbestrol and ethinyl estradiol),antiestrogens (tamoxifen), and androgens (testosterone propionate andfluoxymesterone), radionuclides, toxins and cytotoxic drugs, boronaddends, chemotherapy agents, photodynamic therapy dyes, and antibioticsor combinations thereof to treat cancer. Many toxins and cytotoxic drugsare known in the art that have cytotoxic effects on cells, any of whichcan be used in connection with the methods provided herein. Examples ofknown cytotoxic agents useful in the present methods are listed, forexample, in Goodman et al., “The Pharmacological Basis of Therapeutics,”Sixth Edition, A. G. Gilman et al., eds., Macmillan Publishing Co., NewYork (1980). These include, but are not limited to, adrenocorticalsuppressants, such as mitotane; alkyl sulfonates, such as busulfan;ethylenimine derivatives, such as thiotepa; nitrosoureas, such ascarmustine, lomustine, semustine and streptozocin; folic acid analogs,such as methotrexate; methyl hydrazine derivatives, such asprocarbazine; nitrogen mustards, such as mechlorethamine,cyclophosphamide, melphalan, uracil mustard and chlorambucil; purineanalogs, such as mercaptopurine and thioguanine; pyrimidine analogs,such as fluorouracil, cytarabine and azaribine; substituted ureacompounds, such as hydroxyurea; taxol; triazenes, such as dacarbazine;and vinca alkaloids, such as vinblastine and vincristine.

Any antibiotic known in the art, such as aminoglycosides, bleomycin,cephalosporins and other beta-lactam antibiotics, chloramphenicol,clindamycin, dactinomycin, daunorubicin, doxorubicin, fusidic acid,macrolides, metronidazole, mithramycin, mitomycin, mupirocin,penicillins, rifamycins, sulfonamides, tetracyclines, trimethoprim andbeta-lactam inhibitors, can be included in the formulation. Drugs thatinterfere with intracellular protein synthesis also can be used in themethods provided herein; such drugs are known to those skilled in theart and include puromycin, cycloheximide, and ribonuclease.

The methods provided herein also can include administration of one ormore than one compound of Formula I, II or III or pharmaceuticallyacceptable salts or prodrugs thereof in combination with dyes usedphotodynamic therapy for the treatment of cancer, and used inconjunction with appropriate non-ionizing and ionizing radiation. Theuse of porphyrins and other dyes used in photodynamic therapy can beused in the methods herein. Photodynamic therapy for the treatment ofcancer is well known in the art (e.g., see U.S. Pat. Nos. 7,018,395,7,011,812, 6,806,284, 6,723,750, 6,710,066, 6,630,128 and 6,622,729).

6. Methods of Treating Prostate Cancer

In certain embodiments, provided herein are methods for treating,preventing, suppressing, inhibiting or reducing the incidence ofprostate cancer in a subject. The methods include administering one ormore than one compound of Formula I, II or III or pharmaceuticallyacceptable salts or prodrugs thereof in a therapeutically effectiveamount to treat the cancer. In some embodiments, the prostate cancer isandrogen dependant prostate cancer. In certain embodiments, the prostatecancer is androgen independent prostate cancer. In certain embodiments,the prostate cancer is androgen independent, but androgen receptordependant prostate cancer. In some embodiments, administration of theone or more than one compound provided herein to a subject afflictedwith prostate cancer alleviates the prostate cancer by killing thecancerous cells. In one embodiment, administration of the one or morethan one compound provided herein to a subject afflicted with prostatecancer results in the inhibition of growth and/or metastasis of theprostate cancer. In some embodiments, the one or more than one compoundof Formula I, II or III or pharmaceutically acceptable salts or prodrugsthereof is co-administered with another therapeutic agent, including,but not limited to, flutamide, bicalutamide and nilutamide, anti-tumoragent, such as toxins and cytotoxic drugs, which can be selectivelytargeted to react with prostate tumors by conjugating to a prostatetumor antigen, and radionuclides.

In certain embodiments, methods are provided for delaying theprogression of prostate cancer in a subject suffering from prostatecancer. The methods include administering to the subject one or morethan one compound of Formula I, II or III or pharmaceutically acceptablesalts or prodrugs thereof, in an amount effective to delay theprogression of prostate cancer in the subject.

7. Methods of Contraception

In certain embodiments, provided herein are methods for providingcontraception in a subject. The methods include administering to thesubject one or more than one compound of Formula I, II or III orpharmaceutically acceptable salts or prodrugs thereof, in an amounteffective to provide contraception in the subject. In some embodiments,provided herein are methods for providing contraception in a malesubject. The methods include administering to the subject one or morethan one compound of Formula I, II or III or pharmaceutically acceptablesalts or prodrugs thereof, in an amount effective to suppress spermproduction in the subject, thereby effecting contraception in thesubject. In one embodiment, the compounds provided herein inhibitspermatogenesis in a subject. In one embodiment, the method includesco-administering one or more than one compound of Formula I, II or IIIor pharmaceutically acceptable salts or prodrugs thereof with anandrogen, such as 19-nortestosterone, 7α-methyl-19-nortestosterone and5α-dihydro-testosterone. In one embodiment, the method includesco-administration of a compound of Formula I, II or III orpharmaceutically acceptable salts or prodrugs thereof that is an ARantagonist with testosterone.

8. Methods of Providing Hormone Therapy

In certain embodiments, provided herein are methods for providinghormone therapy to a subject. The method includes administering to thesubject one or more than one compound of Formula I, II or III orpharmaceutically acceptable salts or prodrugs thereof, in an amounteffective to modulate androgen receptor activity, and thereby effect achange in an androgen-dependent condition.

9. Methods of Treating Postmenopausal Conditions

In certain embodiments, provided herein are methods for treating,preventing, suppressing, inhibiting or reducing the incidence ofpostmenopausal conditions in a subject. The methods includeadministering to the subject one or more than one compound of Formula I,II or III or pharmaceutically acceptable salts or prodrugs thereof, inan amount effective to treat the postmenopausal condition. In oneembodiment, the postmenopausal condition treated by the method includes,but is not limited to, loss of libido, decreased sexual activity,diminished feelings of physical well-being, fatigue and hot flashes. Inone embodiment, the method includes co-administering one or more thanone compound of Formula I, II or III or pharmaceutically acceptablesalts or prodrugs thereof with one or more estrogens, such as estrone,2-hydroxyestrone, 2-methoxyestrone, 4-hydroxyestrone,15-α-hydroxyestrone, 16-α-hydroxyestrone, 16-β-hydroxyestrone, estradiol(17β-estradiol), 2-hydroxy-estradiol, 2-methoxy-estradiol,4-hydroxy-estradiol, 16-oxoestradiol, estriol, 16-epiestriol and17-epiestriol or combinations thereof. In one embodiment, the methodincludes co-administering one or more than one compound of Formula I, IIor III or pharmaceutically acceptable salts or prodrugs thereof with oneor more estrogenic compound, such as estradiol valerate, estrone,estrone sulfate, an estrone sulfate piperazine salt or an ester thereof,or a synthetic estrogen. In one embodiment, the method includesco-administering one or more than one compound of Formula I, II or IIIor pharmaceutically acceptable salts or prodrugs thereof with one ormore agents selected from among alendronate, calcitonin, clodronate,clomiphene, clomiphene citrate, clonidine, conjugated estrogen, naturalestrogen, synthetic estrogen, ethinyl estradiol, estradiol,enclomiphene, enclomiphene citrate, etidronate, ibandronate,medroxyprogesterone acetate, megestrol acetate, norethindrone acetate,pamidronate, progesterone, risedronate, tiludronate, zuclomiphene,zuclomiphene citrate and combinations thereof.

10. Methods of Treating Hematopoietic Disorders

Also provided herein are methods of treating, preventing, suppressing,inhibiting or reducing the incidence of a hematopoietic disorder in asubject. The methods include administering to the subject one or morethan one compound of Formula I, II or III or pharmaceutically acceptablesalts or prodrugs thereof, in an amount effective to treat thehematopoietic disorder. In some embodiments, the hematopoietic disorderincludes, but not limited to, anemia, leukemia, and hematopoieticconditions caused by bone marrow transplantation or chemo-/radiationtherapy. Also provided are methods of increasing the number of red bloodcells in a mammal in need thereof. The method includes administering atherapeutically effective amount of one or more than one compound ofFormula I, II or III or pharmaceutically acceptable salts or prodrugsthereof, in an amount effective to increase the number of red bloodcells in a subject. Also provided are methods of treating anemia,thrombocytopenia or neutropenia in a subject. The methods includeadministering to the subject in need of such treatment one or more thanone compound of Formula I, II or III or pharmaceutically acceptablesalts or prodrugs thereof, in an amount effective to treat anemia,thrombocytopenia or neutropenia in the mammal. In some embodiments ofthese methods, one or more than one compound of Formula I, II or III orpharmaceutically acceptable salts or prodrugs thereof is co-administeredwith a therapeutically effective amount of at least one hematopoieticcytokine. In some embodiments, the hematopoietic cytokine is selectedfrom among erythropoietin, granulocyte-colony stimulating factor,granulocyte-macrophage-colony stimulating factor, interleukin-1,interleukin-3, interleukin-4, interleukin-5, interleukin-7,interleukin-9, interleukin-11, macrophage-colony stimulating factor,stem cell factor and thrombopoietin.

Also provided are methods of increasing serum EPO levels in a subject.The methods include administering a therapeutically effective amount ofone or more than one compound of Formula I, II or III orpharmaceutically acceptable salts or prodrugs thereof, in an amounteffective to increase the serum EPO levels in the subject.

11. Methods of Treating Neurodegenerative Diseases and Disorders

In some embodiments, provided herein are methods of treating,preventing, suppressing, inhibiting or reducing the incidence of aneurodegenerative disease or disorder in a subject. The methods includeadministering to a subject having a neurodegenerative disease ordisorder, one or more than one compound of Formula I, II or III orpharmaceutically acceptable salts or prodrugs thereof, in an amounteffective to treat the neurodegenerative disease or disorder. In someembodiments, the neurodegenerative disorder is Alzheimer's disease. Insome embodiments, methods for preventing the onset or delaying theprogression of Alzheimer's disease in patients are provided. The methodincludes administering to a subject one or more than one compound ofFormula I, II or III or pharmaceutically acceptable salts or prodrugsthereof, in an amount effective to prevent the onset or delay theprogression of Alzheimer's disease in a subject. The method can includeco-administering an effective amount of one or more than one compound ofFormula I, II or III or pharmaceutically acceptable salts or prodrugsthereof with a therapeutically-effective amount of a compound thatinhibits the formation or release of β-amyloid.

Any of the known inhibitors of the formation or release of β-amyloid canbe used in the methods, including, but not limited to, compoundsdescribed in U.S. Pat. App. Pub. Nos. U.S. 2002/0025955, 2002/0022621and U.S. 2003/0114496 and in WO 03/018543, WO 01/53255, WO 01/66564, WO01/70677, WO 01/90084, WO 01/77144, WO 02/30912, WO 02/36555, WO02/081435, WO 02/081433, WO 98/28268, WO 02/47671, WO 99/67221, WO01/34639, WO 01/34571, WO 00/07995, WO 00/38618, WO 01/92235, WO01/77086, WO 01/74784, WO 01/74796, WO 01/74783, WO 01/60826, WO01/19797, WO 01/27108, WO 01/27091, WO 00/50391 and WO 02/057252.

12. Methods of Treating Cognitive Impairment

Also provided herein are methods for treating, preventing, suppressing,inhibiting or reducing the incidence of cognitive impairment in asubject. The methods include administering to a subject having cognitiveimpairment one or more than one compound of Formula I, II or III orpharmaceutically acceptable salts or prodrugs thereof, in an amounteffective to treat the cognitive impairment.

13. Methods of Treating Depression

Also provided herein are methods for treating, preventing, suppressing,inhibiting or reducing the incidence of depression in a subject. Themethods include administering to a subject having cognitive impairmentone or more than one compound of Formula I, II or III orpharmaceutically acceptable salts or prodrugs thereof, in an amounteffective to treat depression.

14. Methods of Treating Obesity

Also provided herein are methods of treating, preventing, suppressing,inhibiting or reducing the incidence of obesity in a subject. Themethods include administering to a subject one or more than one compoundof Formula I, II or III or pharmaceutically acceptable salts or prodrugsthereof, in an amount effective to treat obesity. In one embodiment, acompound of Formula I, II or III or pharmaceutically acceptable salts orprodrugs thereof that is an AR agonist is used to treat a male subjectwith abdominal adiposity. In one embodiment, a compound of Formula I, IIor III or pharmaceutically acceptable salts or prodrugs thereof that isan AR antagonist is used to treat a female subject with abdominalobesity.

15. Methods of Treating Insulin Resistance and Diabetes

Provided herein are methods of treating, preventing, suppressing,inhibiting or reducing the incidence of insulin resistance in a subject.The methods include administering to a subject one or more than onecompound of Formula I, II or III or pharmaceutically acceptable salts orprodrugs thereof, in an amount effective to treat insulin resistance.Also provided herein are methods of treating, preventing, suppressing,inhibiting or reducing the incidence of type 2 diabetes in a subject.The methods include administering to a subject one or more than onecompound of Formula I, II or III or pharmaceutically acceptable salts orprodrugs thereof, in an amount effective to treat type 2 diabetes. Insome embodiments, the method for treating diabetes includesco-administering an effective amount of one or more than one compound ofFormula I, II or III or pharmaceutically acceptable salts or prodrugsthereof with an effective amount of an anti-diabetic drug, such as, butnot limited to, thiazolidinedione-type drugs such as pioglitazone orrosiglitazone, sulfonylurea-type drugs, such as chlorpropamide,glimepiride, glipizide, glyburide or tolbutamide, a biguanide-type drugsuch as metformin, exenatide, acarbose, repaglinide, nateglinide,tolazamide or combinations thereof.

Also provided herein are methods of treating, preventing, suppressing,inhibiting or reducing the incidence of arterial hypertension,hyper-insulineamia, hyperglycaemia or dyslipidaemia characteristicallyappearing with insulin resistance. The methods include administering toa subject one or more than one compound of Formula I, II or III orpharmaceutically acceptable salts or prodrugs thereof, in an amounteffective to treat arterial hypertension, hyperinsulinemia,hyperglycaemia, type 2 diabetes or dyslipidaemia characteristicallyappearing with insulin resistance.

16. Methods of Treating Sexual Dysfunction

In certain embodiments, provided herein are methods of treating,preventing, suppressing, inhibiting or reducing the incidence of sexualdysfunction in a subject. The methods include administering to thesubject one or more than one compound of Formula I, II or III orpharmaceutically acceptable salts or prodrugs thereof, in an amounteffective to treat or prevent sexual dysfunction in the subject. In someembodiments, the sexual dysfunction is male erectile dysfunction. Insome embodiments, the sexual dysfunction is impotence.

In certain embodiments, provided herein are methods of increasing thelibido of a male or female subject. The methods include administering tothe subject in need thereof one or more than one compound of Formula I,II or III or pharmaceutically acceptable salts or prodrugs thereof in anamount that is effective to increase the libido of the subject.

17. Methods of Treating Arthritic Conditions and Inflammatory Disorders

Also provided herein are methods for treating, preventing, suppressing,inhibiting or reducing the incidence of an arthritic condition orinflammatory disorder. The methods include administering one or morethan one compound of Formula I, II or III or pharmaceutically acceptablesalts or prodrugs thereof in an amount effective for the treatment orprevention of an arthritic condition or an inflammatory disorder. In oneembodiment, the arthritic condition or inflammatory disorder is selectedfrom among osteoarthritis, Behcet's disease, bursitis, tendonitis, CPPDdeposition disease, carpal tunnel syndrome, Ehlers-Danlos syndrome,fibromyalgia, gout, infectious arthritis, inflammatory bowel disease,juvenile arthritis, lupus erythematosus, Lyme disease, Marfan syndrome,myositis, osteoarthritis, osteogenesis imperfecta, osteonecrosis,polyarteritis, polymyalgia rheumatica, psoriatic arthritis, Raynaud'sphenomenon, reflex sympathetic dystrophy syndrome, Reiter's syndrome,rheumatoid arthritis, scleroderma and Sjogren's syndrome. In oneembodiment, the method is for treating, preventing, suppressing,inhibiting or reducing the incidence of osteoarthritis, which includesadministering one or more than one compound of Formula I, II or III orpharmaceutically acceptable salts or prodrugs thereof in an amounteffective for the treatment or prevention of osteoarthritis. In certainembodiments of these methods, the one or more than one compound ofFormula I, II or III or pharmaceutically acceptable salts or prodrugsthereof is co-administered with one or more drugs or agents known totreat or prevent arthritic conditions, such as corticosteroids,cytotoxic drugs (or other disease modifying or remission inducingdrugs), gold treatment, methotrexate, aspirin, NSAIDs, COX-2 inhibitorsand DMARDs (Disease-Modifying Anti-Rheumatic Drugs).

Exemplary DMARDs include, but are not limited to, leflunomide,auranofin, sulfasalazine, mycophenolate, myochrysine, cyclosporine,cyclophosphamide, azathioprine, chlorambucil, methotrexate, minocycline,penicillamine and hydroxychloroquine. Exemplary NSAIDs include, but arenot limited to, diclofenac/misoprostol, diclofenac potassium,diclofenac, diflunisal, etodolac, fenoprofen, flurbiprofen, ibuprofen,indomethacin, ketoprofen, meclofenamate, mefanamic acid, meloxicam,nabumetone, naproxen and naproxen sodium, oxaprozin, piroxicam, sodiumsulindac and tolmetin. Exemplary COX-2 inhibitors include, but are notlimited to, celecoxib, rofecoxib and valdecoxib.

18. Methods of Improving Lipid Profile

In certain embodiments, provided herein are methods of improving thelipid profile in a subject. The methods include administering to thesubject one or more than one compound of Formula I, II or III orpharmaceutically acceptable salts or prodrugs thereof in an amounteffective to effect the lipid profile in the subject. In one embodiment,the one or more than one compound of Formula I, II or III orpharmaceutically acceptable salts or prodrugs thereof is co-administeredwith another agent, such as an anti-cholesterol agent or lipid-loweringagent, such as, but not limited to, β-hydroxy-β-methylbutyric acid,lactoferrin, cholestyramine, colestipol, colesevelam, nicotinic acid,fibric acids (gemfibrozil, fenofibrate and clofibrate) and HMG-coAreductase inhibitors (lovastatin, pravastatin, simvastatin, fluvastatin,atorvastatin and cerivastatin).

In certain embodiments, provided herein are methods of reducingcirculating lipid levels in a subject. The method includes administeringto the subject one or more than one compound of Formula I, II or III orpharmaceutically acceptable salts or prodrugs thereof in an amounteffective to reduce circulating lipid levels in the subject.

19. Methods of Treating Atherosclerosis

In certain embodiments, provided herein are methods of treating,preventing, suppressing, inhibiting or reducing the incidence ofatherosclerosis and its associated diseases including cardiovasculardisorders, cerebrovascular disorders, peripheral vascular disorders, andintestinal vascular disorders in a subject. The methods includeadministering to the subject one or more than one compound of Formula I,II or III or pharmaceutically acceptable salts or prodrugs thereof,alone or in combination with a selective estrogen receptor modulator(SERM) compound.

20. Methods of Treating Conditions Related to Androgen Decline

In certain embodiments, provided herein are methods of treating,preventing, suppressing, inhibiting or reducing the incidence of acondition related to androgen decline, such as in a male subject. Themethods include administering to the subject one or more than onecompound of Formula I, II or III or pharmaceutically acceptable salts orprodrugs thereof in an amount effective to treat the condition relatedto androgen decline in the subject. In some embodiments, the conditionis selected from among fatigue, depression, decreased libido, sexualdysfunction, erectile dysfunction, hypogonadism, osteoporosis, hairloss, obesity, sarcopenia, osteopenia, benign prostate to hyperplasia,anemia, alterations in mood and cognition, and prostate cancer.

21. Methods of Treating Conditions Related to Androgen Deficiency

In certain embodiments, provided herein are methods of treating,preventing, suppressing, inhibiting or reducing the incidence of acondition related to androgen deficiency, such as in a female subject.The methods include administering to the subject one or more than onecompound of Formula I, II or III or pharmaceutically acceptable salts orprodrugs thereof, in an amount effective to treat the condition relatedto androgen decline in the subject. In one embodiment, the condition isselected from among sexual dysfunction, decreased sexual libido,sarcopenia, osteopenia, osteoporosis, alterations in cognition and mood,depression, anemia, hair loss, obesity, endometriosis, breast cancer,uterine cancer and ovarian cancer.

J. COMBINATION THERAPIES

In certain embodiments, one or more compounds or compositions providedherein can be co-administered with one or more other therapeutic agents.In certain embodiments, such one or more other therapeutic agents aredesigned to treat the same disease or condition as the one or morecompounds or pharmaceutical compositions provided herein. In certainembodiments, such one or more other therapeutic agents are designed totreat a different disease or condition as the one or more compounds orcompositions provided herein. In certain embodiments, such one or moreother therapeutic agents are designed to treat an undesired effect ofone or more compounds or compositions provided herein. In certainembodiments, one or more compounds or compositions provided herein isco-administered with another therapeutic agent to treat an undesiredeffect of that other agent.

In certain embodiments, compounds or compositions provided herein andone or more other therapeutic agents are administered at the same time.In some embodiments, compounds or compositions provided herein and oneor more other therapeutic agents are administered at the differenttimes. In certain embodiments, compounds or compositions provided hereinand one or more other therapeutic agents are prepared together in asingle formulation. In certain embodiments, compounds or compositionsprovided herein and one or more other therapeutic agents are preparedseparately.

Examples of therapeutic agents that can be co-administered withcompounds or compositions provided herein include, but are not limitedto, analgesics (e.g., acetaminophen); anti-inflammatory agents,including, non-steroidal anti-inflammatory drugs (e.g., ibuprofen, COX-1inhibitors and COX-2, inhibitors); salicylates; anti-biotics;antivirals; antifungal agents; antidiabetic agents (e.g., biguanides,glucosidase inhibitors, insulins, sulfonylureas, andthiazolidenediones); adrenergic modifiers; diuretics; hormones (e.g.,anabolic steroids, androgen, estrogen, calcitonin, progestin,somatostan, and thyroid hormones); immuno-modulators; muscle relaxants;anti-histamines; osteoporosis agents (e.g., bisphosphonates, calcitonin,and estrogens); prostaglandins, antineoplastic agents; psychotherapeuticagents; sedatives; antibodies; and vaccines.

In other embodiments, therapeutic agents that can be co-administeredwith compounds or compositions provided herein include, but are notlimited to, other modulators of nuclear hormone receptors or othersuitable therapeutic agents useful in the treatment of theaforementioned disorders including: anti-diabetic agents;anti-osteoporosis agents; anti-obesity agents; anti-inflammatory agents;anti-anxiety agents; anti-depressants; anti-hypertensive agents;anti-platelet agents; anti-thrombotic and thrombolytic agents; cardiacglycosides; cholesterol/lipid lowering agents; mineralo-corticoidreceptor antagonists; phosphodiesterase inhibitors; protein tyrosinekinase inhibitors; thyroid mimetics (including thyroid receptoragonists); anabolic agents; HIV or AIDS therapies; therapies used in thetreatment of Alzheimer's and other cognitive disorders; therapies usedin the treatment of sleeping disorders; anti-proliferative agents;anti-tumor agents; bisphosphonates; estrogens; SERMs; anti-estrogens;cathepsin inhibitors; α_(v)β₃ integrin receptor antagonists; calcitonin;PPARγ inhibitors; osteoprotegerin; and proton pump inhibitors.

K. EXAMPLES

The following examples, including experiments and results achieved, areprovided for illustrative purposes only and are not to be construed aslimiting the claimed subject matter.

Example 1

4-(2(R)-(1(S)-hydroxyl-2,2,2-trifluoroethyl)-pyrrolidinyl)-2-trifluoromethyl-benzonitrile(Compound 101) andR,R-4-(2-(1-Hydroxyl-2,2,2-trifluoroethyl)-pyrrolidinyl)-2-trifluoromethyl-benzonitrile(Compound 102)

A mixture of D-prolinol, 4-fluoro-2-trifluoromethylbenzonitrile, andtriethylamine in THF was stirred over night at 60° C. Standard work-upof the reaction mixture providedR-4-(2-hydroxylmethylpyrrolidinyl)-2-trifluoromethyl-benzonitrile inmoderate yield. The intermediate alcohol was oxidized by sulfur trioxidepyridine complex to giveR-4-(2-formyl-pyrrolidinyl)-2-trifluoromethyl-benzonitrile. The aldehydeintermediate was treated with trimethyl(trifluoromethyl)-silane toprovide a mixture of two diastereomers. HPLC separation generated pureforms of Compounds 101 and 102.

Compound 101: ¹H-NMR (500 MHz, CDCl₃) 7.59 (d, J=8.8, 1H), 7.07 (d,J=2.9, 1H), 6.92 (dd, J=8.8 and 2.9, 1H), 4.24 (t, J=7.5, 1H), 3.91 (t,J=6.3, 1H), 3.63 (dd, J=7.8 and 9.3, 1H), 3.29 (td, J=6.9 and 9.7, 1H),2.55 (s, 1H), and 2.05-2.23 (m, 4H).

Compound 102: ¹H-NMR (500 MHz, acetone-d₆) 7.78 (d, J=8.8, 1H), 7.01 (d,J=2.9, 1H), 6.94 (dd, J=8.8 and 2.9, 1H), 5.68 (bd, J=3.3, 1H),4.44-4.50 (m, 1H), 4.36 (d, J=8.3, 1H), 3.64-3.77 (m, 1H), 3.44 (td,J=9.8 and 7.8, 1H), 2.30-2.47 (m, 2H), and 2.07-2.17 (m, 2H).

Example 2

R, R,R-4-(2-(1-Hydroxyl-2,2,2-trifluoroethyl)-5-methylpyrrolidinyl)-2-trifluoro-methylbenzonitrile(Compound 103) and4-(2(R)-(1(S)-hydroxyl-2,2,2-trifluoroethyl)-5(R)-methylpyrrolidinyl)-2-trifluoromethylbenzonitrile(Compound 104)

Compounds 103 and 104 can be prepared in a similar fashion as describedin Example 1 by using D-pyroglutamic acid as a starting material.

Compound 103: ¹H-NMR (500 MHz, CDCl₃) 7.54 (d, 1H, J=8.5), 6.88 (d, 1H,J=2.3), 6.68 (dd, 1H, J=8.5 and 2.3), 4.41-4.32 (m, 1H), 4.19-4.15 (m,1H), 3.98-3.93 (m, 1H), 2.79 (d, 1H, J=5.6), 2.59-2.49 (m, 1H),2.17-1.98 (m, 2H), 1.93-1.85 (m, 1H), 1.35 (d, 3H, J=6.1).

Compound 104: ¹H-NMR (500 MHz, CDCl₃) 7.62 (d, 1H, J=8.8), 7.16 (d, 1H,J=2.3), 6.99 (dd, 1H, J=8.8 and 2.3), 4.21-4.15 (m, 1H), 3.95-3.84 (m,2H), 2.60 (d, 1H, J=3.5), 2.43-2.34 (m, 1H), 2.04-1.99 (m, 2H),1.94-1.72 (m, 1H), 1.40 (d, 3H, J=6.1).

Example 3

R,R-4-(2-(1-Hydroxyl-2,2,2-trifluoroethyl)pyrrolidinyl)-2-chlorobenzonitrile(Compound 105) and4-(2(R)-(1(S)-hydroxyl-2,2,2-trifluoroethyl)pyrrolidinyl)-2-chlorobenzonitrile(Compound 106)

Compounds 105 and 106 can be prepared in a similar fashion as describedin Example 1 by using 2-chloro-4-fluorobenzonitrile as a startingmaterial.

Example 4

R, R,R-4-(2-(1-Hydroxyl-2,2,2-trifluoroethyl)-5-methylpyrrolidinyl)-2-chlorobenzonitrile(Compound 107) and4-(2(R)-(1(S)-hydroxyl-2,2,2-trifluoroethyl)-5(R)-methylpyrrolidinyl)-2-chlorobenzonitrile(Compound 108)

Compounds 107 and 108 can be prepared in a similar fashion as describedin Example 1 by using 2-chloro-4-fluorobenzonitrile as a startingmaterial.

Compound 107: ¹H-NMR (500 MHz, CDCl₃) 7.39 (d, 1H, J=8.8), 6.61 (d, 1H,J=2.3), 6.46 (dd, 1H, J=8.8 and 2.3), 4.41-4.36 (m, 1H), 4.14-4.09 (m,1H), 3.93-3.88 (m, 1H), 2.78 (d, 1H, J=5.6), 2.55-2.49 (m, 1H),2.13-1.98 (m, 2H), 1.88-1.81 (m, 1H), 1.33 (d, 3H, J=6.4).

Compound 108: ¹H-NMR (500 MHz, CDCl₃) 7.45 (d, 1H, J=9.1), 6.89 (d, 1H,J=2.3), 6.76 (dd, 1H, J=9.1 and 2.3), 4.15-4.09 (m, 1H), 3.89-3.80 (m,2H), 2.66 (d, 1H, J=2.6), 2.41-2.31 (m, 1H), 2.03-1.96 (m, 2H),1.82-1.69 (m, 1H), 1.38 (d, 3H, J=6.2).

Example 5

R,R-4-(2-(1-Hydroxyl-2,2,2-trifluoroethyl)pyrrolidinyl)-2-chloro-3-methyl-benzonitrile(Compound 109) and4-(2(R)-(1(S)-hydroxyl-2,2,2-trifluoroethyl)-pyrrolidinyl)-2-chloro-3-methylbenzonitrile(Compound 110)

Compounds 109 and 110 can be prepared in a similar fashion as describedin Example 1 by using 2-chloro-4-fluoro-3-methylbenzonitrile as astarting material.

Compound 109: ¹H-NMR (500 MHz, CDCl₃) 7.43 (d, 1H, J=8.5), 6.92 (d, 1H,J=8.5), 4.19-4.14 (m, 1H), 4.00-3.95 (m, 1H), 3.71-3.63 (m, 1H),3.02-2.95 (m, 1H), 2.39 (d, 1H, J=4.4), 2.34 (s, 3H), 2.32-2.21 (m, 1H),2.17-2.00 (m, 2H), 1.93-1.80 (m, 1H).

Compound 110: ¹H-NMR (500 MHz, CDCl₃) 7.45 (d, 1H, J=8.5), 7.10 (d, 1H,J=8.5), 4.30-4.25 (m, 1H), 3.80-3.78 (m, 1H), 3.61-3.53 (m, 1H), 3.10(bs, 1H), 2.91-2.84 (m, 1H), 2.40 (s, 3H), 2.43-2.32 (m, 1H), 2.14-1.90(m, 2H), 1.89-1.81 (m, 1H).

Example 6 Co-Transfection Assay—AR Agonist/Antagonist Activity

The ability of Compound 102(4-(2(R)-(1(R)-hydroxyl-2,2,2-trifluoroethyl)-pyrrolidinyl)-2-trifluoromethylbenzonitrile)to activate or repress the ability of AR to induce gene expression wasassessed using the cotransfection assay.

CV-1 cells (African green monkey kidney fibroblasts) were cultured inthe presence of Dulbecco's Modified Eagle Medium (DMEM) supplementedwith 10% charcoal resin-stripped fetal bovine serum then transferred to96-well microtiter plates one day prior to transfection.

The CV-1 cells were transiently transfected by calcium phosphatecoprecipitation according to the procedure of Berger et al., J. SteroidBiochem. Mol. Biol. 41: 733 (1992) with the following plasmids: pShAR (5ng/well), MTV-LUC reporter (100 ng/well), pRS-β-Gal (50 ng/well) andfiller DNA (pGEM; 45 ng/well). The receptor plasmid, pRShAR, containsthe human AR under constitutive control of the SV-40 promoter, as morefully described in J. A. Simental et al., “Transcriptional activationand nuclear targeting signals of the human androgen receptor”, J. Biol.Chem. 266: 510 (1991). The reporter plasmid, MTV-LUC, contains the cDNAfor firefly luciferase (LUC) under control of the mouse mammary tumorvirus (MTV) long terminal repeat, a conditional promoter containing anandrogen response element. See e.g., Berger et al. supra. In addition,pRS-β-Gal, coding for constitutive expression of E. coli β-galactosidase(β-Gal), was included as an internal control for evaluation oftransfection efficiency and compound toxicity.

Six hours after transfection, media was removed and the cells werewashed with phosphate-buffered saline (PBS). Media containing referencecompounds (e.g., progesterone as a PR agonist, mifepristone((11β,17β)-11-[4-(dimethylamino)-phenyl]-17-hydroxy-17-(1-propynyl)estra-4,9-dien-3-one:RU486; Roussel Uclaf) as a PR antagonist; dihydrotestosterone (DHT;Sigma Chemical) as an AR agonist and 2-OH-flutamide (the activemetabolite of2-methyl-N-[4-nitro-3-(trifluoromethyl)-phenyl]-pronanamide;Schering-Plough) as an AR antagonist; estradiol (Sigma) as an ER agonistand ICI-164,384(N-butyl-3,17-dihydroxy-N-methyl-(7-alpha,17-beta)-estra-1,3,5(10)-triene-7-undecanamide;ICI Americas) as an ER antagonist; dexamethasone (Sigma) as a GR agonistand RU486 as a GR antagonist; and aldosterone (Sigma) as an MR agonistand spironolactone((7-alpha-[acetylthio]-17-alpha-hydroxy-3-oxopregn-4-ene-21-carboxylicacid gamma-lactone; Sigma) as an MR antagonist) and/or Compound 102 inconcentrations ranging from 10⁻¹² to 10⁻⁵ M were added to the cells.Three to four replicates were used for each sample. Transfections andsubsequent procedures were performed on a Biomek 1000 automatedlaboratory work station.

After 40 hours, the cells were washed with PBS, lysed with a TritonX-100-based buffer and assayed for LUC and 13-Gal activities using aluminometer or spectrophotometer, respectively. For each replicate, thenormalized response (NR) was calculated as:

$\frac{{LUC}\mspace{14mu}{response}}{\beta\text{-}{Gal}\mspace{14mu}{rate}}\mspace{14mu}$where   β-Gal  rate = β-Gal ⋅ 1 × 10⁻⁵/β-Gal  incubation  time.

The mean and standard error of the mean (SEM) of the NR were calculated.Data was plotted as the response of the compound compared to thereference compounds over the range of the dose-response curve. Foragonist experiments, the effective concentration that produced 50% ofthe maximum response (EC₅₀) was quantified. Agonist efficacy was afunction (%) of LUC expression relative to the maximum LUC production bythe reference agonist for PR, AR, ER, GR or MR. Antagonist activity wasdetermined by testing the amount of LUC expression in the presence of afixed amount of DHT as an AR agonist and progesterone as a PR agonist atthe EC₅₀ concentration. The concentration of test compound thatinhibited 50% of LUC expression induced by the reference agonist werequantified (IC₅₀). In addition, the efficacy of antagonists wasdetermined as a function (%) of maximal inhibition.

Compound 102 produced a concentration-dependent increase in luciferaseactivity in MDA-MB-453 cells that have endogenous expression of hAR.Compound 102 did not produce a concentration-dependent decrease inluciferase activity when tested in the presence of an EC₅₀ concentrationof DHT (1 nM). Compound 102 was found to be a potent activator of hARwith an EC₅₀ of 3.1 nM and an efficacy of 59% when expressed relative todihydrotestosterone (DHT, EC₅₀ of 2 nM and an efficacy of 100%), anatural ligand for AR. The ability of Compound 102 to activatetranscription by other nuclear receptors also was evaluated. Agonistefficacy was determined relative to the appropriate reference agonistfor each nuclear receptor. Compound 102 did not transactivate(demonstrate agonist activity) through the other nuclear receptorsevaluated (hPR-B, hGR, hMR, hERα, hRARα, hLXRα, hFRα, hPPARα, hPPARγ,hPPARδ), except it very weakly activated hRXRα. Compound 102 antagonizeddexamethasone-induced activation of hGR (60% with an IC₅₀ of 1.6 μM) andprogesterone-induced activation of hPR-B (74% with IC₅₀ of 280 nM), butnot activation of hERα or hMR.

Example 7 Receptor Binding Assays

Preparation of Receptor Proteins

A baculovirus expression plasmid including cDNA encoding the humanandrogen receptor (hAR) was prepared using standard techniques. Theexpression plasmid was used to transfect Spodopter frugiperda-21 (Sf-21)cells. A suspension culture of uninfected Sf-21 cells was grown to adensity of 1.2×10⁶ cells/mL and then infected with the recombinant virusincluding hAR cDNA at a multiplicity of infection of 2. The infectedSf-21 cells were incubated for 48 hours and then collected bycentrifugation at 1000×g for 10 minutes at 4° C. The resulting cellpellets were resuspended in lysis buffer (50 mM potassium phosphatebuffer, pH 7.0, 10 mM monothioglycerol, 5 mM DTT, 20 mM sodiummolybdate, 1 mM PMSF, 1 μg/mL aprotinin, and 10 μg/mL leupeptin) andincubated for 15 minutes on ice. The resuspended cell pellets werehomogenized using a Dounce homogenizer and a B pestle. A volume of 2 MKCl was added to the homogenized cell pellets to a final concentrationof 0.4 M. The resulting receptor lysates were centrifuged at 100,000×gfor 60 min at 4° C. and stored for use in binding assays.

Binding Assays

Binding assay samples were prepared in separate mini-tubes in a 96-wellformat at 4° C. Each binding assay sample was prepared in a volume of250 μL of Receptor-Assay Buffer (10% glycerol, 25 mM sodium phosphate,10 mM potassium fluoride, 10 mM sodium molybdate, 0.25 mM CHAPS, 2 mMDTT and 1 mM EDTA, (adjusted to pH 7.5)) containing 50 μg of receptorlysate; 2-4 nM of [³H]-dihydro-testosterone at 50-100 Ci/mmol; andeither a reference compound or a test compound. Each reference compoundand test compound was assayed at varying concentrations, ranging from3.2×10⁻¹⁰ to 10⁻⁵ M. Each concentration of each reference compound andeach test compound was assayed in triplicate. The assay samples wereincubated for 16 hours at 4° C.

After incubation, 200 μL of 6.25% hydroxylapatite in assay buffer wasadded to each assay sample to precipitate the protein. The assay sampleswere then centrifuged and the supernatants were discarded. The resultingpellets were washed twice with assay buffer lacking DTT. Radioactivityin counts per minute (CPM) of each washed pellet was determined byliquid scintillation counter (MicroBeta™, Wallach).

After correcting for nonspecific binding, IC₅₀ values were determinedusing a 4-parameter fit, such as provided in commercially availablesoftware for curve fitting (e.g., Xlfit curve fitting software, IDBSScientific Products, Guildford, UK). Typically all 4 parameters werefloated to allow the best fit to converge. The K_(i) values weredetermined by application of the Cheng-Prusoff equation to the IC₅₀values using previously determined K_(d) values for each specificligand.

Compound 102 and other androgen modulating compounds, includingR,R-2,2,2-trifluoro-1-[1-(4-nitro-3-trifluoromethyl-phenyl)-pyrrolidin-2-yl]-ethanol(LG0893410) andS,R-2,2,2-trifluoro-1-[1-(4-nitro-3-trifluoromethyl-phenyl)-pyrrolidin-2-yl]-ethanol(LG0893411):

were tested for their ability to bind to hAR. Compound 102 demonstratedconsistently higher potency, with a K_(i) of 0.9 nM, by 3-6 fold thanLG0893410 (K_(i) of 2.5 nM) and LG0893411 (K_(i) of 5.7 nM) for bindingto baculovirus expressed hAR protein. Higher potency indicates thatCompound 102 can be administered at a lower dose to achieve equaltherapeutic effect.

Example 8 Activation of hAR-Responsive Luciferase Reporter

A functional assay was conducted to determine the ability of testsubstances to modulate gene expression via endogenous human androgenreceptor (hAR) in a human cell line. A luciferase reporter assay wasutilized to establish the ability of test compounds to activatehAR-regulated gene expression in a human cell line containingendogenously expressed hAR (MDAMB-453 cells) transfected with anandrogen-responsive luciferase reporter plasmid containing the cDNA forfirefly luciferase. MDA-MB-453 cells were transfected with a plasmidcontaining the cDNA for firefly luciferase (LUC) under the control of aconditional promoter containing hormone response elements recognized byAR (MMTV-LUC) and a P-galactosidase expression plasmid (pRS-(3-Gal) fornormalization of transfection efficiency. The reporter plasmid MMTVLUCcontains the mouse Mammary Tumor Virus (MMTV) Long Terminal Repeat (LTR)conditional promoter containing hormone response elements recognized byhAR. The luciferase response was calculated as:Luciferase response/β-gal ratewhere

Luciferase response=Relative Luciferase Units (RLU) and

β-gal rate=β-gal 0.D.₄₁₅/β-Gal incubation time in minutes.

The mean of the normalized response at each concentration of compoundwas calculated. The effective concentration that produced 50% of themaximum response (EC₅₀) was determined for each compound byinterpolation between two concentrations spanning the midpoint of theconcentration-response curve. Agonist efficacy for test compounds wascalculated as a percent of normalized response relative to the maximumnormalized response by the reference agonist DHT.

Each of Compound 102, LG0893410 and LG0893411 activated AR-dependentgene expression. Compound 102, having an EC₅₀ of 2.8 nM, was 8.5 to 12.5times more potent in activating hAR-responsive luciferase reporter thanLG0893410 (EC₅₀ of 24.2 nM) and LG0893411 (EC₅₀ of 35.0 nM). Higherpotency indicates that Compound 102 can be administered at a lower doseto achieve equal therapeutic effect.

Example 9 Activation of hAR in a Co-Transfection Assay

A luciferase reporter co-transfection assay also was used to evaluatethe ability of compounds provided herein to activate hAR regulated geneexpression. Compound 102 and other androgen receptor modulators,including LG0893410 and LG0893411, were assayed to assess their abilityto activate hAR regulated gene expression by transfecting CV-1 cells,derived from African monkey kidney and lacking endogenous AR, with anexpression plasmid containing the cDNA for the human AR (pRShAR) and theandrogen-responsive luciferase reporter plasmid (MMTV-LUC). CV-1 cellswere transiently transfected with pRShAR, a P-galactosidase expressionplasmid (pRS-(3-Gal) for normalization of transfection efficiency, andthe reporter plasmid MMTV-LUC using a non-liposomal formulation, theFuGENE 6 transfection reagent.

The mean of the normalized response at each concentration of compoundwas calculated. The EC₅₀ was determined for each compound and agonistefficacy for test compounds was calculated as a percent of normalizedresponse relative to the maximum normalized response by the referenceagonist DHT.

Each of Compound 102, LG0893410 and LG0893411 activated hAR in theco-transfection assay. Compound 102, having an EC₅₀ of 4.4 nM, was 3 to5 times more potent in activating hAR than LG0893410 (EC₅₀ of 13.0 nM)and LG0893411 (EC₅₀ of 23.3 nM). Higher potency indicates that Compound102 can be administered at a lower dose to achieve equal therapeuticeffect.

Example 10 Skeletal α-Actin Promoter Assay

Compound 102 was tested to determine its ability to activate humanskeletal α-actin using the skeletal α-actin promoter assay. A mousemuscle cell line (C2C12) was transiently transfected with a human ARexpression plasmid and a luciferase reporter plasmid containing theskeletal α-actin promoter upstream of the luciferase cDNA. C2C12 cells(at a concentration of 5×10⁵) were seeded in a T25 flask and culturedfor 24 hours. The cells were harvested by using trypsin and seeded at6000 cells/well in 96-well plates. The same day, a non-liposomalformulation, FuGENE 6 reagent (Roche, Indianapolis, Ind.) was used fortransfection of the cells. To each well was added 45.5 ng of theskeletal α-actin-LUC reporter plasmid, 4.55 ng pRShAR and 5 ng pRS-BG ascarrier DNA. Twenty-four hours after transfection, differentconcentrations of the solvated compound (10⁻¹⁰ to 10⁻⁶ M) were added intriplicate to the cells and the cells were incubated for 24 hours. Themedium was aspirated and the cells were lysed with adetergent-containing buffer. After addition of Luciferase Assay Bufferto each well, the cells were assayed for luciferase activity using aluminometer to determine the level of transcriptional activation. Theeffective concentration that produced 50% of the maximum response (EC₅₀)was determined from the concentration-response curve for the compound byinterpolation between two concentrations spanning the midpoint of theconcentration response curve. Agonist efficacy for the test compound wascalculated as a percent of luciferase response relative to the maximumluciferase response by the reference agonist DHT.

Compound 102 stimulated skeletal α-actin promoter activity in aconcentration-dependent manner. At a concentration of 1×10⁻⁷ M, Compound102 increased the promoter activity about 17.5-fold compared to vehiclecontrol. Compound 102 was not as potent as DHT but was equallyefficacious. Compound 102 activated the skeletal α-actin promoter withan average EC₅₀ of 8.7 nM (DHT 1 nM) and efficacy of 93% of DHT.Compound 102 showed a strong AR-mediated stimulation of skeletal α-actinpromoter activity in muscle cells, indicating that this compound likelyup-regulated skeletal α-actin production at the transcription level.Compound 102 stimulates α-actin promoter activity in muscle cells in amanner similar to that of DHT, is highly potent (8.7 nM) and efficacious(93% compared to DHT).

The ability of Compound 102 to stimulate muscle cells was compared toother androgen modulating compounds, including LG0893410 and LG0893411.Compound 102 was found to be 5-10 fold more potent in stimulating musclecells than LG0893410 and LG0893411, having an EC₅₀ of 8.7 nM compared to45.2 nM for LG0893410 and 85.6 nM for LG0893411. Compound 102 also wasnearly as effective as the androgen agonist DHT in stimulating musclecells (93% of DHT), while LG0893410 and LG0893411 were much lesseffective (having an efficacy of 43% and 59%, respectively) instimulating α-actin gene transcription in muscle cells.

Example 11 IL-6 Promoter Repression Assay

IL-6 is an important bone resorption factor. Over-expression of IL-6 cancause severe bone loss in vivo. Reducing IL-6 production by bone cellswould be therapeutically beneficial by reducing bone resorption.Compound 102 and the androgen receptor modulators LG0893410 andLG0893411 were tested to determine their ability to suppressTNFα-IL-1β-induced IL-6 promoter activity in Saos-2 human osteoblastcells using a cotransfection assay. A human osteoblast cell line(Saos-2) was transiently transfected with a human AR expression plasmidand a luciferase reporter plasmid containing the IL-6 promoter upstreamof the luciferase cDNA. Saos-2 cells were seeded at 6000 cells/well in96-well plates. The same day, a non-liposomal formulation, FuGENE 6reagent (Roche, Indianapolis, Ind.) was used for transfection of thecells. To each well was added 3.6 μg of the IL-6-LUC reporter plasmidand 0.7 μg of pCMV-hAR. Twenty-four hours after transfection, differentconcentrations of the solvated compound (10⁻¹¹ to 10⁻⁶ M) in DMEM(Dulbecco's Minimal Essential Media) were added in triplicate to thecells and the cells were incubated for 24 hours. The expression ofluciferase was stimulated by the cytokines Tumor Necrosis Factor α(TNFα, 10 ng/mL) and Interleukin-1 beta (IL-1β, 1 ng/mL). The medium wasaspirated and the cells were lysed with a detergent-containing buffer.After addition of Luciferase Assay Buffer to each well, the cells wereassayed for luciferase activity using a luminometer to determine thelevel of transcriptional activation. The effective concentration thatproduced 50% of the maximum response (EC₅₀) was determined from theconcentration-response curve for the compound by interpolation betweentwo concentrations spanning the midpoint of the concentration responsecurve. Agonist efficacy for the test compound was calculated as apercent of luciferase response relative to the maximum luciferaseresponse by the reference agonist DHT.

All three test substances were full agonists compared to DHT insuppressing IL-6 production by human bone cells. The addition ofCompound 102 in concentrations from 10⁻¹¹ to 10⁻⁶ M resulted in aconcentration-dependent suppression of IL-6 promoter activity to thelevel seen in the absence of TNFα and IL-1β. Compound 102 displayedpotent and efficacious AR-mediated suppression of TNFα-IL-1β-inducedIL-6 promoter activity in Saos-2 human osteoblast cells. Compound 102was found to be highly potent, having an average IC₅₀ of 0.41 nM (DHThad an average IC₅₀ of 0.03 nM, LG0893410 had an average IC₅₀ of 1.20 nMand LG0893411 had an average IC₅₀ of 1.50 nM) and Compound 102 was foundto have an efficacy of 97% of DHT. Compound 102 suppresses IL-6 promoteractivity in a manner similar to that of DHT, and is 3-4 fold more potentthan LG0893410 and LG0893411. These results suggest that Compound 102down-regulates IL-6 production at the transcriptional level. BecauseCompound 102 suppresses IL-6 production by osteoblasts, it is expectedto have a beneficial in vivo effect on bone by reducing bone resorption.

Example 12 Metabolic Stability in Liver Microsomes

Liver microsomes are subcellular fractions that containdrug-metabolizing enzymes, such as cytochrome P450, flavinmonooxygenases and UDP glucuronyl transferases. When the elimination ofa drug occurs primarily by metabolism, the route of metabolism cansignificantly affect the drug's safety and efficacy, hence affecting thedirections for use of the drug. Rat liver microsomes are a commonly usedanimal model for determination of drug metabolism (e.g., seeShayeganpour et al., Drug Metab Dispos 34 (1): 43-50 (2006)).

The metabolism of Compound 102 was evaluated in rat liver microsomes.The metabolism was initiated by adding 25 μL of cofactor (100 mM ofglucose 6-phosphate, 20 mM of NADP, 20 mM of glucose-6-phosphatedehydrogenase, and 63 mM of UDPGA) to wells containing 50 μL of 5 mg/mLof rat liver microsomes and 425 μL, of 0.59 mM of Compound 102 made in100 mM phosphate buffer (pH 7.4) at 37° C. At 0, 5, 10 and 20 minutes, a60 μL aliquot of the sample was withdrawn. The unmetabolized testsubstance remaining in the sample was determined using HPLC and massspectrometry and the half-life for degradation of the compound wascalculated in minutes using curve fitting. For Compound 102, thehalf-life was determined to be 44.5 minutes (average value determined in2 experiments with each experiment using triplicate measurements),demonstrating that Compound 102 has high metabolic stability, since itis only slowly metabolized.

High metabolic stability promotes oral bioavailability and reduces theburden of xenobiotic metabolites generated after administration of atherapeutically effective dose. High metabolic stability is preferredfor drugs that are intended for systemic administration, such as wouldbe the case for treating diseases or conditions such as aplastic anemia;anorexia; arthritis, including inflammatory arthritis, rheumatoidarthritis, osteoarthritis and gout; arteriosclerosis; atherosclerosis;bone disease, including metastatic bone disease; bone damage orfracture, such as by accelerating bone fracture repair and/orstimulation of osteoblasts and/or stimulation of bone remodeling and/orstimulation of cartilage growth; distraction osteogenesis; reduced bonemass, density or growth; bone weakening, such as induced byglucocorticoid administration; musculoskeletal impairment (e.g., in theelderly); cachexia; cancer, including breast cancer and osteosarcoma;cardiac dysfunction (e.g., associated with valvular disease, myocardialinfarction, cardiac hypertrophy or congestive heart failure);cardiomyopathy; catabolic side effects of glucocorticoids; Crohn'sdisease; growth retardation in connection with Crohn's disease; shortbowel syndrome; irritable bowel syndrome; inflammatory bowel disease;ulcerative colitis; cognitive decline and impairment; dementia; shortterm memory loss; chronic obstructive pulmonary disease (COPD); chronicbronchitis; decreased pulmonary function; emphysema; decreased libido inboth men and women; depression; nervousness, irritability and/or stress;reduced mental energy and low self-esteem (e.g.,motivation/assertiveness); dyslipidemia; erectile dysfunction; frailty;age-related functional decline (“ARFD”) in the elderly; growth hormonedeficiency; hematopoietic disorders; hormone replacement (male andfemale); hypercholesterolemia; hyperinsulinemia; hyperlipidemia;hypertension; hyperandrogenemia; hypogonadism (including primary andsecondary); hypothermia (including hypothermia following anesthesia);impotence; insulin resistance; type 2 diabetes; lipodystrophy (includingin subjects taking HIV or AIDS therapies such as protease inhibitors);male menopause; metabolic syndrome (syndrome X); loss of muscle strengthand/or function (e.g., in the elderly); muscular dystrophies; muscleloss following surgery (e.g., post-surgical rehabilitation); muscularatrophy (e.g., due to physical inactivity, bed rest or reducedweight-bearing conditions such as microgravity); neuro-degenerativediseases; neuromuscular disease; decreased platelet count; plateletaggregation disorders; obesity; osteoporosis; osteopenia;glucocorticoid-induced osteoporosis; osteochondro-dysplasias;periodontal disease; premenstrual syndrome; postmenopausal symptoms inwomen; Reaven's syndrome; rheumatological disease; sarcopenia; male andfemale sexual dysfunction (e.g., erectile dysfunction, decreased sexdrive, sexual well-being, decreased libido); physiological shortstature, including growth hormone deficient children and short statureassociated with chronic illness and growth retardation associated withobesity; tooth damage (such as by acceleration of tooth repair orgrowth); thrombocytopenia; vaginal dryness; atrophic vaginitis;ventricular dysfunction; wasting, including wasting secondary tofractures and wasting in connection with chronic obstructive pulmonarydisease (COPD), chronic liver disease, AIDS, weightlessness, cancercachexia, burn and trauma recovery, chronic catabolic state (e.g.,coma), eating disorders (e.g., anorexia), chemotherapy, multiplesclerosis or other neurodegenerative disorders.

Example 13 In Vivo Assay—Orchidectomized Mature Male Rats

The orchidectomized male Sprague-Dawley rat model was used to assess theeffects of Compound 102 on various reproductive, gonadotropin andmusculoskeletal endpoints, including sex accessory organs, bone, serumgonadotropin levels and striated muscle in mature male rats.

In this assay, two-month old rats were acclimated in a vivarium for oneweek. After this acclimation period, rats were surgicallyorchidectomized under isoflurane anesthesia and left untreated for 14days. After 14 days, animals were sorted into groups such that nostatistically significant differences in mean body weights wereobserved. Rats began receiving treatment 14 days after the day ofsurgery. Sham-operated and orchidectomized rats, treated with vehicle,served as controls. The different test groups were treated with variousdoses of Compound 102 (0.03 to 100 mg/kg/day per os (by mouth)). Dosagesincluded 0.03, 0.1, 0.3, 1, 3, 10, 30 and 100 mg/kg/day. Ostarine, anon-steroidal androgenic drug (Evans et al., J Clin Oncology 25: 9119(2007)) was dosed at dosages similar to other test groups as acomparison. After the 14^(th) dose, venous blood was collected at 0, 1,2, 4 and 6 hours after dosing. The blood samples were collected inEDTA-containing tubes (Becton Dickinson, Franklin Lakes, N.J.).Approximately 24 hours after the last dose, animals were euthanized andseminal vesicle weights, ventral prostate weights, levator ani weights,preputial gland weights and blood samples were collected on necropsy.

A. Serum and Plasma Collection

For serum, whole blood (5 mL) was collected in vacutainer tubes (BectonDickinson) and the blood samples were allowed to clot at roomtemperature for about 2 hours. Serum was separated by centrifugation at2500 rpm for 30 minutes, collected and frozen (−80° C.). For plasma,whole blood was collected in EDTA-containing tubes by centrifugation at1000×g for 5 minutes at 4° C. Plasma samples were stored at −20° C.

B. Sample Preparation

For the preparation of all dose formulations, the appropriate amount oftest compound (Compound 102) for the highest concentration to beadministered was weighed. The compound was suspended in the formulationvehicle (5% Tween®-80 polysorbate, 90% PEG-400 polyethylene glycol and5% PVP-K30 polyvinyl-pyrrolidone). The suspension was sonicated for 15minutes, and the resulting formulation was diluted using vehicle toobtain the proper volume and concentrations of the dosing materials toachieve a dosing volume of 1 mL/kg body weight.

C. Luteinizing Hormone (LH) Radioimmunoassay

Serum samples were assayed with a double anti-serum procedure usingreagents from the National Institute of Diabetes and Digestive andKidney Diseases (NIDDK). Sample compounds and standards (NIDDK-rLH-RP-3)in a total volume of 200 μL were incubated at room temperature for 2 to3 days with 100 μL primary anti-serum (rat LH antiserum: rabbitNIDDK-anti-rLH-S-11) diluted 1:100,000. Then, 100 μL of ¹²⁵I-labeled LH(Covance Laboratories Madison, Wis.) diluted 200,000-300,000 cpm/mL wasadded to the tubes and incubation continued for an additional 24 hourperiod.

Bound hormone was separated from free hormone by precipitation with aspecific goat ant-rabbit serum (GARS from Antibodies, Inc., Davis,Calif.). An aliquot of 50 μL of 4% normal rabbit serum was added to eachincubation tube, after which an additional 50 μL of a 1:10 GARS solutionwas added. The tubes were vortexed and incubated overnight at 4° C.

The assay was terminated by centrifugation at 2,500 rpm for 30 minutesin a Sorvall RC 3C Plus centrifuge (rotor #H2000B) at 4° C. Thesupernatants were decanted and discarded and the pellets were counted ina 10-channel gamma-counter.

D. Osteocalcin Immunoradiometric Assay (IRMA)

Serum osteocalcin, a vitamin K-dependent, calcium binding protein, is abiochemical marker that reflects bone formation activity (Gundberg, JClin Ligand Assay 21: 128-138 (1998)). Serum osteocalcin was quantitatedusing an IRMA kit from Immutopics, Inc. (#50-1500, San Clemente,Calif.). The procedures were performed following the protocols suppliedby the vendor (Rat Osteocalcin IRMA Kit Cat. #50-1500, 2005) except that25 μL of sample was used instead of 5 μL of sample as suggested. Thetest uses two different antibodies to measure the serum osteocalcin. Thesample is incubated simultaneously with a bead-immobilized goat antibodythat recognizes the mid-region C-terminal portion of the osteocalcinmolecule and an ¹²⁵I-labeled goat antibody that recognizes the aminoterminal portion of the molecule. Osteocalcin present in the samplebinds to the antibody immobilized on the bead and the radiolabeledantibody binds to the osteocalcin.

For sample dilution, 25 μL of sample and 400 μL of zero standard weredispensed into appropriately labeled tubes and vortexed. A 100 μLaliquot of standard, diluted control or diluted sample was dispensedinto appropriately labeled tubes. 200 μL of ¹²⁵I-labeled Rat OsteocalcinAntibody was dispensed into all tubes followed by vortex mixing. Onebead including the goat antibody was added to each tube, tilting thetube to prevent splashing of the tube contents by the addition of thebead. The tubes were sealed with Parafilm® and incubated at roomtemperature for 18 to 24 hours. After the incubation period, thecontents of each tube was aspirated, leaving the bead in the tube. Thebead in each tube was washed three times by dispensing 2 mL of washsolution into each tube and then completely removing the wash solutionby aspiration. Each tube was counted in a gamma counter for one minuteand the counts recorded. A standard curve was generated using the ratosteocalcin standards provided in the kit. The rat osteocalcinconcentration (in ng/mL) of the diluted controls and diluted sampleswere read directly from the standard curve.

E. Quantitation of Compound 102 in Plasma Samples

The determination of plasma concentration was done by an LC-MS/MSmethod. The standard solution of each drug was spiked in blank ratplasma, calibration standards were constructed from 0.0001 μg/mL to 10μg/mL; and 50 μL of each calibration standard was extracted with 250 μlof acetonitrile containing 10 ng/mL of internal standard in a 96-wellplate. Also, 50 μL of plasma sample was extracted with 250 μL ofacetonitrile containing internal standard in a 96-well plate.

The Q1/Q3 mass was 337.1/267.3 amu for Compound 102 in a negative modeand 338.0/269.1 amu for ostarine in a negative mode.

F. Pharmacokinetic Analysis

Plasma concentration time data for each animal were analyzed usingWinNonlin (version 5.0, Pharsight WinNolin copyright © 1998-2005) bycon-compartmental PK methods (Gibaldi et al., Pharmokinetics (2^(nd)ed., Marcel Dekker, New York, N.Y., pp. 271-318 and 409-417 (1982)). Theelimination half life (t_(1/2)) was not estimated in this study due tothe fact that plasma samples were collected at four time points (1, 2, 4and 6 hours postdose). The area under the plasma concentration curve(AUC) was calculated by the trapezoidal method. The 0 hour plasmaconcentration was considered as the 24 hour plasma concentration at Day14, assuming steady state, and used to estimate AUC₂₄. Peakpharmacokinetic (PK) parameters were calculated and rounded to threedecimal places.

G. Data Analysis

Results were analyzed by analysis of transformed data (Box et al, J RoySoc Series B 26: 211-252 (1964) and Box et al., Technometrics 16:385-389 (1974). When necessary, transformations were performed to ensurethat variances were homogeneous among groups and that the residuals ofthe one-way analysis of variance model followed a Gaussian (normal)distribution. When the analysis of variance reached significance, datawere further evaluated by the Fisher's LSD test. A P value lee than 0.05was considered as the minimum criterion to declare statisticallysignificant differences. When possible, data were fitted to a modifiedfour-parameter logistic equation (Ghosh et al., J Biopharm Stat 8:645-665 (1998). The model estimates ED₅₀s in a logarithmic scale, sincelog ED₅₀ is a more robust estimate of the potency. The model alsoprovides an SE for the estimate that is used to calculate 95% confidencelimits of the estimated potencies. The following equation exemplifiesthe basic, reparametrized four-parameter logistic equation used in thesestudies:Y=(A−D)/(1+e ^(B)·(log C−log x))+Dwhere A is the maximum, D is the minimum, B is the slope factor, C iseither the EC₅₀ of the IC₅₀, depending on the direction of the responseand x is the dose of the compound used.

Results

The 14-day rat model is a short term in vivo model used to demonstratethe selectivity of androgen modulator compounds. Orchidectomy removesalmost all of the endogenous circulating androgens in the rat. WhenCompound 102 was orally administered at levels of about 1.5 mg/kg/day,Compound 102 was able to maintain levator ani weight at sham-equivalentlevels, while the same dose did not maintain the growth of ventralprostate or seminal vesicle at sham-equivalent level. At the highesttested dose (100 mg/kg/day), Compound 102 significantly increasedlevator ani weight above sham-equivalent levels, but still only restoredthe ventral prostate or seminal vesicles to approximately 50% of shamlevels. These findings suggest that Compound 102 when administeredorally can sustain levator ani weight similar to or greater than intactcounterparts while maintaining androgen-sensitive sex accessory glandsat weights lower than intact male rats.

Compound 102 had an inhibitory effect on serum LH and was able tomaintain sham-equivalent levels at a dose of 10 mg/kg/day. Compound 102significantly decreased serum LH below sham equivalent levels at thehighest tested dose (100 mg/kg/day).

Preputial gland weight, measured as a marker of sebaceous glandactivity, remained below vehicle equivalent levels for all doses ofCompound 102 less than 10 mg/kg/day PO. Compound 102 increased preputialgland weight above the sham level at the 30 mg/kg/day dose. The effectwas statistically significant at this dose.

Serum osteocalcin tended to decrease with increasing dose of Compound102, but the effects were not statistically significant at any testeddose. Orchidectomy has been reported to increase serum osteocalcin inmale rats 2 weeks post-surgery (Ivaska et al., J Biol Chem 279:18361-18369 (2004)).

The test compound was quantified in most of the plasma samplescollected, although Compound 102 was below the limit of quantitation(BLQ) in a few plasma samples at 0.03 mg/kg/day and 0.1 mg/kg/day doses.The mean PK parameters for Compound 102 are shown below in Table 1:

TABLE 1 Pharmacokinetic Parameters at Day 14 (Mean ± SD). 0.03 0.01 0.31 3 10 30 100 Parameter mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kgAUC₆ 0.002 ± 0.004 ± 0.018 ± 0.063 ± 0.113 ± 0.370 ± 0.645 ± 0.998 ± (μg· hr/ml) 0.001 0.005 0.008 0.037 0.069 0.141 0.406 0.480 AUC₂₄ * 0.005 ±0.015 ± 0.042 ± 0.123 ± 0.234 ± 0.764 ± 1.881 ± 4.053 ± (μg · hr/ml)0.007 0.012 0.029 0.063 0.140 0.315 1.120 1.987 C_(MAX) 0.001 ± 0.002 ±0.012 ± 0.018 ± 0.036 ± 0.111 ± 0.205 ± 0.321 ± (μg/ml) 0.001 0.0010.012 0.010 0.019 0.050 0.169 0.112 t_(MAX) (hr)  2.0 ±  2.8 ±  1.3 ± 2.5 ±  1.3 ±  1.8 ±  4.3 ±  3.5 ± 1.6 2.2 1.0 1.7 0.5 1.5 2.4 2.9 * The0 hr plasma concentration was substituted for the 24 hr plasmaconcentration in order to estimate AUC₂₄.The systemic exposures, as measured by AUC₂₄ and C_(MAX), increased asthe dose level increased and were roughly dose proportional. The peakplasma concentrations were observed between 1 hour and 5 hour postdose.

Pharmacokinetic analysis indicated that exposure of Compound 102increased with dose up to 100 mg/kg/day PO. Expressed relative toexposure (AUC₂₄), levator ani and ventral prostate weights increasedwith Compound 102 exposure in a dose dependent manner. Compound 102increased levator ani to sham equivalent levels at an exposure ofapproximately 0.2 μg·/mL. At exposures of 1 μg·/mL or greater, Compound102 stimulated muscle mass above sham equivalent levels yet restored theventral prostate to less than 50% of sham equivalent levels. Ostarinewas not as potent as Compound 102, restoring levator ani weight to shamequivalent levels at an exposure of 4 μg·/mL. At exposures greater than10 μg·/mL, ostarine stimulated muscle mass above sham levels but did notrestore ventral prostate to sham equivalents at any dose.

Thus, in orchidectomized male rats, Compound 102 exhibited positiveactivity in musculoskeletal endpoints. Compound 102 had doseproportional exposure and maintained levator ani weight atsham-equivalent levels at a dose of 2 mg/kg/day (AUC₂₄: 0.2 μg·/mL). Atdoses that maintained levator ani weights at sham equivalent levels,Compound 102 had lower efficacy (relative to sham) in sex accessory andsebaceous glands. In this model, Compound 102 showed tissue selectiveactivity toward the muscle endpoint. Compound 102 showed lower potencyfor reducing the elevated LH levels in castrate rats (≧10 mg/kg/day)compared to increasing the reduced levator ani skeletal muscle mass inthese animals (2 mg/kg/day). Compound 102 suppressed serum LH belowsham-equivalent levels at high doses. The comparator compound, ostarine(a known non-steroidal selective androgen receptor modulator (SARM)),had similar efficacy relative to Compound 102 and also demonstrated hightissue selectivity.

The finding that Compound 102 increases the levator ani skeletal musclein rats with abnormally low muscle mass suggests that the compound canbe used for treating sarcopenia in humans (e.g., see Joseph et al., MolAspects Med 26: 181-201 (2005)). Based on the ≈2 mg/kg/day dose in ratsfor restoration of skeletal muscle mass (equivalent to ≈14 mg/m² bodysurface area), the dose for therapeutic effect in humans would targetsystemic exposure of about 0.1-0.5 μg·hr/mL, such as 0.2 μg·hr/mL inpatients with sarcopenia.

Example 14 In Vivo Assay—Ovariectomized Mature Female Rats

In animal models of osteoporosis, androgens increase bone density andstrength in male rats (Wakely et al., J Bone & Mineral Research 6:325-330 (1991) and Vandenput et al., Calcified Tissue International 70:170-175 (2002)) and female rats (Tobias et al., Am J Physiology 267:E853-E859 (1994) and Coxam et al., Bone 19: 107-114 (1996)). Theovariectomized mature female rat model was used to assess the effects oforally administered compounds provided herein, such as Compound 102, onbone as well as other efficacy and side-effect endpoints includingmuscle mass, fat mass and clitoral gland weight.

In this assay, approximately three-month old rats were acclimated in avivarium for one week. After acclimation, rats were anesthetized withAvertin and the ovariectomy (OVX) was performed using the dorsalapproach. For the sham procedure, the surgical procedure was conductedper protocol, and the ovaries exteriorized but not removed. Afterrecovery, no further experimental manipulations were performed for sevenweeks, at which point the rats were scanned by dual energy x-rayabsorptiometry and sorted into experimental groups based upon femoralbone mineral content (BMC). Three dosages of Compound 102 were tested(0.03, 0.3 and 3 mg/kg/day PO). For comparison, ostarine, a knownnon-steroidal SARM, was tested at the same dosages (0.03, 0.3 and 3mg/kg/day PO). In addition, reference compounds estradiol andtestosterone were administered. Estradiol was administeredsubcutaneously to one ovariectomized group at a dose of 0.1 mg/kg/dayand testosterone propionate was administered subcutaneously to anotherovariectomized group at a dose of 1 mg/kg/day.

Treatment continued once daily for 12 weeks, at which point animals wereeuthanized and tissues were collected for further analysis. Animalsreceived subcutaneous injections of fluorochrome markers for bonehistomorphometric analysis. Alizarian Red S was prepared as a 3%solution and given at a dose of 30 mg/kg at the time of the baselinescan. Calcein was administered 10 days and 3 days prior to necropsy.

After the 14^(th) dose, approximately 250 μL venous blood was collectedvia percutaneous jugular puncture from the rats at 0, 0.5, 1, 2, 4 and 6hours post-dosing. The blood samples were collected into lithium-heparintubes (Becton Dickinson, Franklin Lakes, N.J.). Plasma fractions werecollected for pharmacokinetic analysis. Approximately 24 hours after thelast dose, rats were sacrificed by cardiac exsanguination, and blood wascollected into serum separator tubes (Becton Dickinson), and thegastrocnemius muscle, plantaris muscle, uterus, clitoral glands,clitoris and inguinal fat pads were isolated, blotted and weighedindividually. Uterine weight measurements can be inaccurate insurgically ovariectomized rats due to the potential loss of uterinetissue at the tips of the uterine horns during removal of the ovaries.To minimize this variability, the length of the uterine horns wasmeasured and the uterine weight was reported normalized to uterinelength. The left femur and lumbar vertebra L5 were wrapped insaline-soaked gauze and stored at −20° C. for biochemical analysis. Theright femur and lumbar vertebrae L3-L4 were collected and stored in 70%ethanol for histomorphometry. The right tibia was collected in formalinfor histological analysis. The left tibia was collected on dry ice andstored at −20° C. for measurement of alkaline phosphatase activity.

A. Sample Preparation

Compound 102 and ostarine individually were formulated in a solutioncontaining 5% Tween-80 polysorbate, 90% PEG-400 polyethylene glycol, and5% polyvinyl-pyrrolidone K30. The mixture was stirred and sonicated in awater bath for 30 minutes until the compound dissolved into a solution.After sonication, 1% carboxymethyl cellulose (CMC) in water was added tothe solution in a 9:1 ratio (9 parts CMC/water solution to 1 partPEG-400/Tween-80/PVP). Compound 102 and ostarine were administeredorally in a volume of 4 mL/kg.

Reference compounds estradiol and testosterone propionate wereformulated in a vehicle containing 70% polyethylene glycol and 30% DMSO(dimethyl sulfoxide) by volume. Compounds were weighed and added to theappropriate volume of vehicle. The mixture was sonicated for severalminutes to ensure that the compounds were thoroughly dissolved. Thereference compounds were administered subcutaneously in a volume of 0.4mL/kg.

B. Dual Energy X-Ray Absorptiometry (DEXA)

Dual X-ray absorptivity (DEXA) was performed on a Norland PeripheralDual Energy X-Ray Absorptiometer (pDEXA Sabre, Norland Medical Systems,Orthmetrix, White Plains, N.Y.). Prior to each use, the machine wascalibrated using two phantoms supplied by the manufacturer. The machinecalculated a coefficient of variation over the prior 16 calibrations todetermine the measurement precision. The coefficient of variation forthe quality control phantoms was between 0.47% to 0.57% during thecourse of the study.

For in vivo DEXA measurements, the rats were anesthetized withisoflurane (Hospira, Inc., Lake Forrest, Ill.) and placed prone on thebed of the scanner. A scan that encompassed the lumbar spine and theright femur was performed and collected. Regional analysis wereperformed on the whole femur, the mid-diaphyseal femoral shaft and thelumbar spine. The mid-femur measurement included 3.5 mm on either sideof the mid-length of the femur and included the entire cortical width.The lumbar spine was defined as L3-L5 as defined by the intervertebraldisk and included the entire spine width. The bone mineral content(BMC), projected bone area and the areal bone mineral density (BMD) weremeasured for each region, described above. BMD was calculated as the BMCdivided by projected bone area. The coefficient of variations onrepeated scans on live rats was determined to by less than 1.6% formeasurements of the BMC and 2.5% for measurements of the BMD for thefemur and vertebral bodies.

BMC, BMD and bone length were measured by pDEXA on an entire femur afterit was removed from the animal at the end of the experiment. Thecoefficients of variation for the phantom standards were as describedabove for the in vivo measurements. The coefficient of variation forrepeated measurement of the isolated femur was less than 0.4% for theBMC and the BMD measurements.

C. Osteocalcin IRMA

Serum osteocalcin was quantitated using a kit from Immutopics (#50-1500,San Clemente, Calif.), using the procedures described above in Example13.

D. Alkaline Phosphatase Derived from the Tibial Periosteum

The whole, intact tibias were manually cleaned of muscle and otheradherent soft tissue. Whole tibias were enzymatically digested with 0.2%SigmaBlend collagenase Type F (Sigma-Aldrich) for 1 hour at 37° C. Theperiosteal isolate was then sonicated for 20 seconds at 30 A with aKontes Micro Ultrasonic Cell Disrupter Model #KT50 (Vineland, N.J.).Total alkaline phosphatase activity of the homogenate was quantified onan ACE Clinical Chemistry Analyzer (Alfa Wassermann, Fairfield, N.J.)following the manufacturer's protocols.

E. Biomechanical Testing

All biomechanical testing was performed on a QTest2/L materials testingsystem with a 2 kN load cell (MTS, Eden Prairie, Minn.). Data wascollected at 10 Hz from the load cell and the crosshead displacement andanalyzed using software designed for materials testing (TestWorks 4,MTS).

Whole femurs were collected for biomechanical analysis and thawed atroom temperature prior to testing. Femurs were kept moist with salinethroughout the preparation and testing. Total femoral length wasmeasured with handheld digital calipers (Mitutoyo, Japan) and themidpoint of the femur was identified. The medial-lateral andanterior-posterior diameters were measured at the mid-femur withcalipers. The whole femur was placed on a custom-designed 3-pointbending fixture, consisting of stainless steel pins, 0.63 mm indiameter, with a span of 15 mm between the lower supports. A 5N preloadwas applied to the femurs at the mid-femur prior to the start oftesting. Testing was performed with a constant crosshead displacementrate of 20 mm/minute until failure, which was defined as a 50% reductionin load. After failure, cortical thickness was measured at each quadrantof the mid-femur using handheld digital calipers. Peak load and peakdisplacement were measured directly from the load cell and thecrosshead, respectively. Stiffness was calculated from the slope of thelinear region of the load/displacement curve. Energy absorption(EnergyToPeak) was calculated as the area under the load/displacementcurve. All of the measurements listed above describe structural or organlevel properties. In an effort to distinguish overall structural andgeometric changes from intrinsic tissue level changes, additionalendpoints were calculated. These endpoints represent each of the abovedescribed endpoints normalized to the tissue volume and geometry. Pealstress (σ), strain (ε), modulus of elasticity (Young's modulus—E) andmodulus of toughness were calculated using standard equations (e.g., seeTurner et al., Bone 14: 595-608 (1993) and Gere et al., Mechanics ofMaterials (PWS-Kent, Boston (1984)).

Lumbar vertebra L5 were thawed at room temperature prior to testing andwere kept moist with saline throughout the preparation and mechanicaltesting. The lateral and dorsal spinous processes were removed with ahandheld grinding/cutting tool (Dremel, Mount Prospect, Ill.) and thecranial end of the vertebral body was mounted on an aluminum slab with acyanoacrylate glue. The stub was mounted in the grips of a low speed saw(Isomet, Buehler Ltd., Lake Bluff, Ill.) and two plane-parallel sectionwere cut at 4 mm intervals, effectively removing the growth plates andprimary spongiosa from the ends of the vertebral body. The resultantcylinder of bone was measured with handheld digital calipers in thecranial/caudal, medial/lateral and dorsal/ventral directions. Specimenswere placed on the loading platen and a compressive load was applied at20 mm/minute in the cranial/caudal direction until failure of thevertebral body. Peak load and peak displacement were measured directlyfrom the load cell and the crosshead, respectively. Stiffness wascalculated from the slope of the linear region of the load/displacementcurve. Energy absorption was calculated as the area under theload/displacement curve. As with the femur, the organ level propertiesdescribed above were normalized to yield intrinsic of materialproperties of the lumbar spine using standard engineering equations.

F. Bone Histomorphometry

The lumbar vertebral bodies were trimmed, dehydrated and embedded inpolymethyl methacrylate. The femur were trimmed into thirds (proximal,mid-shaft and distal). All were dehydrated in ethanol and the distal andmid-shaft were embedded in polymethyl methacrylate. The proximal portionof the femur was dried and stored. Section of the vertebral bodies andfemur mid-shaft were cut with precision bone saws, the section mountedon plastic slides and the sections were ground to about 30-40 μm inthickness using a bone grinding system. The sections were polished andthe fluoro-chrome makers were viewed in the unstained section. Thesection from the lumbar vertebral bodies were used for histomorphometricindices of cancellous bone, while the sections of the femur mid-shaftswere used for histomorphometric indices of cortical bone.

Indices of cortical bone were measured at the mid-diaphyseal shaft ofthe femur. Measurements were made on a BioQuant Nova Prime imageanalysis system interface with a fluorescence microscope. Allmeasurements were made using the calcein labels. These labels were givennear the end of the study. The following measurements were made:cortical area, marrow area, periosteal and endocortical surfaceperimeter, periosteal and endocortical percent double labeled surface (%dLS), periosteal and endocortical percent single-labeled surface (%sLS), periosteal and endocortical percent mineralization surface (% MS),periosteal and endocortical new bone area, periosteal and endocorticalmineral appositional rate (MAR), periosteal and endocortical boneformation rate, surface referent (BFRs), volume referent (BFRv) andendocortical eroded surface (% ES). Indices of cancellous bone structurewere measured in the lumbar vertebral bodies. The measures were made ina central region that was bordered by the primary spongiosas at bothends. The indices of cancellous bone structure included: bone area,percent bone or trabecular bone volume (%), bone perimeter, bonesurface/volume ration and trabecular thickness.

G. Pharmacokinetic Analysis

Pharmacokinetic analysis was performed as described in Example 13.

H. Data Analysis

Results were analyzed by analysis of transformed or ultratransformeddata (Box et al., J Roy Statist Soc Series B 26: 211-252 (1964) and Boxet al., Technometrics 16: 385-389 (1974). When necessary,transformations were performed to ensure that variances were homogeneousamong groups, and that the residuals of the one-way analysis of variancemodel followed a Gaussian (normal) distribution. When the analysis ofvariance reached significance, data were further evaluated by theFisher's LSD test. A P value less than 0.05 was considered as theminimum criterion to declare statistically significant differences.

i. Body and Organ Weights

Compound 102 significantly increased body weight, gastrocnemius weightand plantaris weight, indicating anabolic activity on skeletal muscle.At the dose tested, testosterone tended to increase body weight gainmore than vehicle, but this increase did not achieve statisticalsignificance. Compound 102 significantly increased weight gain frombaseline compared to vehicle at all the doses tested. Ostarinesignificantly increased weight gain compared to vehicle at the mid andhigh dose levels, but at the low dose there was no significantdifference from vehicle. At the maximally effective does for Compound102 (0.3 mg/kg) and ostarine (0.3 mg/kg), the increased change in bodyweight was similar (mean change of +66 grams and +65 grams,respectively). The increase in body weight resulted from an increase inlean tissue mass rather than in increase in fat mass. Inguinal fat padweight was used as a representative fat pad to assess the effects ofdrug on body fat. Inguinal fat pad weight was increased by ovariectomy(OVX+vehicle vs. Sham, P<0.01) and treatment with either estradiol ortestosterone reduced the elevated fat pad weight due to ovariectomy.However, neither Compound 102 nor ostarine significantly affectedinguinal fat pad weight at any dose. The weights of two skeletalmuscles, gastrocnemius and plantaris, were measured to assess theeffects of drugs on lean body mass. Compound 102 had equal or greaterefficacy on muscle endpoints in comparison to testosterone or ostarine.At the maximally effective dose for each drug (0.3 mg/kg Compound 102 orostarine), the gastrocnemius muscle mass was increased by 17% and 10% byCompound 102 and ostarine, respectively. At the dose administered inthis experiment, testosterone tended to increase muscle weight, but theincrease did not achieve statistical significance for eithergastrocnemius or plantaris muscles.

Clitoral gland weight was measured to assess the effect of drugs onsebaceous glands. Testosterone significantly increased clitoral glandweight at the dose tested. Neither Compound 102 nor ostarine affectedclitoral gland weight at the low dose tested. Both Compound 102 andostarine increased clitoral gland weight at the high dose tested. At themid dose (0.3 mg/kg), which was maximally effective for increasing bodyweight and skeletal muscle mass, Compound 102 and ostarine had similaractivity which was only slightly above the sham level. The relativelysmall increase versus sham was statistically significant (P<0.05) formid dose ostarine whereas Compound 102 did not quite achieve statisticalsignificance versus sham at the mid dose (P>0.05).

Clitoral weight was measured to assess the virilizing effects of drugs.Testosterone had a large impact on clitoral weight in spite of its lackof activity on muscle. Compound 102 and ostarine did not affect clitoralweight at the low dose tested. Both Compound 102 and ostarinesignificantly increased clitoral weight at the high dose, but to aslightly lesser extent than testosterone (P>0.05 testosterone verus highdose Compound 102 or ostarine). At mid dose (0.3 mg/kg), which wasmaximally effective for increasing body weight and skeletal mass,Compound 102 had significantly less activity versus testosterone(P<0.01) whereas ostarine was not significantly different fromtestosterone (P>0.05).

Uterine weight measurements can be inaccurate in surgicallyovariectomized rats due to the potential loss of uterine tissue at thetips of the uterine horns during removal of the ovaries. To minimizethis variability uterine weight was normalized to uterine length. BothCompound 102 and ostarine increased uterine weight to length ratio aboveestrogen deficient OVX controls in a dose dependent manner. Uterineweight to length ratio was significantly (P<0.01) less than shamoperated controls at both the low and mid dose of Compound 102. Uterineweight to length ratio was significantly (P<0.01) less than shamoperated controls for low dose ostarine but not for mid dose ostarine(P>0.05). Compound 102 increased uterine weight to length ratiosignificantly less than ostarine (P<0.01) at both the low and mid dose.At the high dose (3 mg/kg) uterine weight to length ratio wassignificantly (P<0.01) increased above sham levels for Compound 102 andostarine but the compounds were not significantly different from eachother (P>0.05 high dose ostarine versus high dose Compound 102).

ii. Dual Energy X-Ray Absorptiometry

Compound 102 significantly increased bone mineral density (BMD) and bonemineral content (BMC) at the lumbar spine after 12 weeks of dosing. Atthe 0.3 mg/kg dose, Compound 102 was the most efficacious treatment inthis experiment. Testosterone, in particular, did not significantlyincrease lumbar spine BMD or BMC. Estradiol increased spinal bonedensity but did not alter bone mineral content, which is consistent witha different mechanism of action. Ostarine had similar activity relativeto Compound 102. Compound 102 also was effective at the femur, a sitecomprised of both cortical and cancellous bone. Compound 102significantly increased femur BMD and BMC and was the most efficaciouscompound tested in this experiment. Ostarine had similar activity asCompound 102, while testosterone was less efficacious and did notsignificantly increase BMD or BMC above vehicle levels. Similar to theresults at the lumbar spine, estradiol significantly increased femur BMDbut not BMC.

iii. Biochemical Assays

Serum osteocalcin, a marker for bone turnover, was significantlysuppressed by Compound 102 and ostarine in a dose-dependent manner.Estradiol and testosterone also suppressed osteocalcin below the levelsof vehicle-treated OVX controls. Alkaline phosphatase activity fromisolated tibiae was significantly increased by Compound 102 and ostarinetreatment. In contrast, estradiol significantly suppressed tibialalkaline phosphatase activity. Testosterone tended to increase alkalinephosphatase activity, but the differences were not significantlysignificant.

iv. Biomechanical Testing

Compound 102 significantly increased whole femur bending strength andbending stiffness. Ostarine had similar activity, increasing femurbending strength slightly less effectively than Compound 102 butincreasing bending stiffness slightly more effectively than Compound102. The difference between Compound 102 and ostarine were notsignificantly significant at equivalent doses. Testosterone increasedbending stiffness but did not significantly increase bending strength.Estradiol did not have a significant effect on biomechanical propertiesat the femur.

Compression testing of the lumbar spine revealed very minor differencesamong treatment groups. None of the treatments significantly increasedpeak stress or stiffness compared to vehicle controls. These findingsare inconsistent with the DEXA and histomorphometry data. Previousexperiments have consistently demonstrated a correlation between DEXA,histomorphometry and biomechanics. Furthermore, in multiple studiesusing this experimental paradigm, significant effect of estradiol onlumbar spine compressive strength and stiffness was observed. The lackof activity with estradiol in the present experiment is anomalous.

v. Histomorphometry

A greater proportion of bone formation was double-labeled withfluorochrome markers as opposed to single-labeled, indicating a morerapid deposition of new bone. Mineral apposition rates were similarlyincreased by Compound 102. The periosteal surface of the mid-femur is asite with negligible bone resorption rats of this age; hence theincrease in bone formation indicates anabolic activity. Ostarine andtestosterone displayed similar anabolic activity on the periostealsurface, although ostarine, but not testosterone, increased corticalbone area. This is consistent with the DEXA and biomechanical testingdata. Estradiol, in contrast, suppressed bone formation rate andcortical bone cross sectional area. There was minimal effect of anycompound on bone histomorphometry at the endosteal surface in thisexperiment. There was a tendency for decreased endosteal mineralapposition rate (MAR) and bone formation rate; however, there wasinsufficient double-labeled surface to provide quantitative data forstatistical analysis. No statistical comparisons were made for theseendpoints due to the absence of double-labeled surface throughout theexperimental groups.

At a trabecular bone site (lumbar spine), Compound 102 significantlysuppressed mineralizing surface and bone formation rates in adoes-dependent manner. This decrease indicates a suppression of boneremodeling consistent with changes in serum osteocalcin. The net effectwas an increase in trabecular bone volume with the 0.3 mg/kg dose ofCompound 102. Ostarine and estradiol had similar activity on trabecularbone remodeling and bone volume. Testosterone had qualitatively similareffects, but the changes were small and only achieved statisticalsignificance for the mineralizing surface measurement.

vi. Pharmacokinetic Parameters

The mean PK parameters for Compound 102 and the comparator compoundostarine are shown below in Table 2:

TABLE 2 Pharmacokinetic Parameters at Day 14 (Mean ± SD of n = 4) AUC₆AUC₂₄* C_(MAX) t_(MAX) Compound (mg/kg) (μg · hr/ml ± SD) (μg · hr/ml ±SD) (μg/ml ± SD) (hr ± SD) Compound 102 (0.03) 0.056 ± 0.056 0.119 ±0.090 0.039 ± 0.028 3.5 ± 1.9 Compound 102 (0.3) 0.051 ± 0.041 0.229 ±0.289 0.040 ± 0.034 0.5 ± 0.4 Compound 102 (3) 0.404 ± 0.199 0.880 ±0.326 0.207 ± 0.087 0.5 ± 0.0 Ostarine (0.03) 0.125 ± 0.022 0.552 ±0.118 0.035 ± 0.007 2.6 ± 2.4 Ostarine (0.3) 1.083 ± 0.105 4.138 ± 0.4480.332 ± 0.040 0.6 ± 0.2 Ostarine (3) 5.043 ± 1.301 20.644 ± 3.645  1.476± 0.429 1.1 ± 0.6 *The 0 hr plasma concentration was substituted for the24 hr plasma concentration in order to estimate AUC₂₄.The systemic exposure, as measured by AUC₆, AUC₂₄ and C_(MAX), increasedas the dose level increased. Ostarine showed significantly higherexposure that Compound 102. Overall, the peal plasma concentrations ofeach test article were observed between 0.5 hours and 3.5 hour postdose.

Conclusions

Compound 102 is a selective androgen receptor modulator that hasbeneficial effects on bone and muscle in a model of postmenopausalosteoporosis. Compound 102 significantly increased cortical bone mass,density, strength, stiffness and periosteal bone formation rates whenorally administered to osteopenic female rats. These changes demonstratethat Compound 102 has anabolic activity at cortical bone sites and areinconsistent with a compound that predominately inhibits resorption,such as estradiol. Compound 102 suppressed cancellous bone turnoverwhile increasing trabecular bone volume and increasing bone mineraldensity at the lumbar spine. In addition to the effects on bone,Compound 102 increased gastrocnemius muscle weight, plantaris muscleweight and body weight without affecting inguinal fat pad weight.Compound 102 demonstrated some tissue selectivity, as it was moreefficacious on muscle and bone at the 0.3 mg/kg dose than testosterone,yet has reduced activity on the clitoral gland, clitoris or uterusrelative to testosterone. The effects of Compound 102 on bone and musclewere similar to the comparator compound ostarine, although Compound 102had increased potency, reaching maximal efficacy at an exposuresubstantially lower than that of ostarine. The data indicate thatCompound 102 has beneficial effects in an animal model of osteoporosis.

Example 17 Pharmacokinetics of Compound 102 in Sprague Dawley Rats

The pharmacokinetics of Compound 102 following repeat oraladministration for 15 days was evaluated in Sprague-Dawley rats. A totalof 30 rats (15 males and 15 females) were divided into three dose groups(5 males/5 females). Compound 102 was suspended in a vehicle of 2% Tween90: 0.5% carboxymethyl cellulose (high viscosity) (50:50, v/v) atconcentrations of 0.25, 0.75 and 2.5 mg/mL and animals were dosed viaoral gavage at 1, 3 and 10 mg/kg once daily for 15 days. Tap water and adiet of rodent chow were provided to the animals ad libitum.

Blood samples were collected on Day 1 and Day 15. Approximately 0.25 mLof blood was collected from each animal via jugular cannula on Day 1 andfrom tail vein on Day 15. Blood samples were taken 1, 2, 4, 8, 12 and 24hr postdose. Each blood sample was collected into tubes containinglithium heparin and kept on wet ice pending centrifugation (max 2 hrs).Samples were centrifuged under refrigeration (2°-8° C. at 3000 g) for 10minutes. Plasma was transferred into a pre-labeled tube (approximately125 μL) placed on dry ice and stored frozen at −70° C. until analysis.

50 μL of plasma was extracted with 250 μL of acetonitrile and the plasmaconcentration of Compound 102 was determined by an LC-MS/MS method. Theplasma concentration time data were analyzed by a non-compartmentalpharmacokinetic method using WinNonlin. The results for pharmacokinetics(Mean±SD, n=5) is shown in Table 3 (Day 1) and Table 4 (Day 15).

TABLE 3 Pharmacokinetic Parameters on Day 1. 1 mg/kg 3 mg/kg 10 mg/kgParameter Male Female Male Female Male Female AUC₂₄ 0.104 ± 0.413 ±0.318 ± 1.209 ± 1.288 ± 3.965 ± (μg · hr/ml) 0.010 0.042 0.053 0.3910.452 0.334 AUC_(inf) 1.109 ± 0.436 ± 0.341 ± 1.256 ± 1.301 ± 4.206 ±(μg · hr/ml) 0.012 0.036 0.080 0.406 0.459 0.384 C_(MAX) 0.014 ± 0.041 ±0.051 ± 0.129 ± 0.176 ± 0.366 ± (μg/ml) 0.005 0.006 0.009 0.054 0.0690.052 t_(max) (hours)  1.2 ±  1.4 ±  1.2 ±  2.6 ±  1.2 ±  1.4 ± 0.4 0.50.4 1.3 0.4 0.5 t_(1/2) (hours)  5.2 ±  5.4 ±  4.6 ±  4.8 ±  3.5 ±  5.6± 0.8 1.2 2.1 0.7 0.3 0.7

TABLE 4 Pharmacokinetic Parameters on Day 15. 1 mg/kg 3 mg/kg 10 mg/kgParameter Male Female Male Female Male Female AUC₂₄ 0.288 ± 0.428 ±0.876 ± 1.003 ± 2.623³ 4.285 ± (μg · hr/ml) 0.076¹ 0.179 0.141 0.464²0.632¹ AUC_(inf) 0.297 ± 0.443 ± 0.893 ± 1.156 ± 2.668³ 4.535 ± (μg ·hr/ml) 0.082¹ 0.186 0.148 0.504² 0.504¹ C_(MAX) 0.052 ± 0.045 ± 0.133 ±0.139 ± 0.345³ 0.389 ± (μg/ml) 0.021¹ 0.015 0.020 0.035² 0.089¹ t_(max)(hours)  1.3 ±  1.8 ±  1.2 ±  1.8 ± 1.5³  2.0 ± 0.5¹ 1.3 0.4 0.5² 0.0¹t_(1/2) (hours)  4.6 ±  4.6 ±  4.1 ±  3.7 ± 3.9³  5.4 ± 0.4¹ 0.2 0.50.8² 1.5¹ ¹n = 4: 1 animal death due to gavage error. ²n = 3: 2 animaldeaths due to gavage error. ³n = 2: 3 animal deaths due to gavage error.

Compound 102 showed dose proportional increase of systemic exposure at1, 3 and 10 mg/kg dose levels. Female rats showed higher systemicexposure than male rats on Day 1 and Day 15. The systemic exposure ofCompound 102 did not decrease with repeat administration. The systemicexposure in male rats was higher on Day 15 than on Day 1, while that infemale rats remained similar between the two days.

Conclusions

The repeat administration of Compound 102 at pharmacologic doses (1, 3and 10 mg/kg) for 15 days resulted in dose proportional increase ofsystemic exposure and similar (females) or higher (males) systemicexposure compared to single dose administration. At equal doses,systemic exposure of Compound 102 was higher in females than males.

Example 18 Oral Toxicity and Toxicokinetic Study in Sprague Dawley Rats

A Sprague Dawley Rat model was used to study oral toxicity andtoxicokinetics. Animals were assigned to groups by a stratifiedrandomization scheme designed to achieve similar group mean body weightsand the groups were randomly assigned to a dosing to provide control ofbias. A total of 154 Sprague-Dawley rats were assigned to dose groups asshown in Table 5 below.

TABLE 5 Dose groups. No. of No. of Total per Dose level Group TestMaterial Males Females Group (mg/kg/day) Main Study Group 1 VehicleControl 10 10 20 0 2 Low 10 10 20 10 3 Low-Mid 10 10 20 100 4 High-Mid10 10 20 300 5 High 10 10 20 700 Toxicokinetic Groups 6 Vehicle Control3 3 6 0 7 Low 6 6 12 10 8 Low-Mid 6 6 12 100 9 High-Mid 6 6 12 300 10High 6 6 12 700

All animals were dosed via oral gavage once daily for 14 days. Theanimals were evaluated for changes in clinical signs(mortality/morbidity observations, twice daily), cage side observations(general appearance and behavior), food consumption (quantitativelymeasured by weighing), body weights, clinical pathology indicesincluding serum chemistry, hematology, coagulation and urinalysis, andother parameters. Blood samples were collected for toxicokineticsanalysis from Groups 6-10 at various time points on Days 1 and 14 (Group6 4 hours post dose; Groups 7-10 at 1, 4 and 12 hours or 2, 8 and 24hours). The animals were fasted for at least 8 hours prior to bloodcollections for serum chemistry and urine collection. Serum chemistryparameters tested included alanine aminotransferase, total protein,aspartate aminotransferase, albumin, alkaline phosphatase, globulin,gamma-glutamyl-transferase, albumin/globulin ratio, lactatedehydrogenase, glucose, total bilirubin, cholesterol, urea nitrogen,triglycerides, creatinine, sodium, calcium, potassium, phosphorus andchloride. Hematology parameters analyzed included red blood cell count,reticulocyte count, hemoglobin concentration, red blood celldistribution width, mean corpuscular volume, mean platelet volume, meancorpuscular hemoglobin concentration, white blood cell count anddifferentials and mean corpuscular hemoglobin. Urinalysis parametersincluded color/character, ketones, specific gravity, bilirubin, pH,occult blood, protein, glucose and microscopics. Main study animals wereeuthanized on Day 15. At termination, a full necropsy was conducted andall tissues were collected, preserved, processes and examinedmicroscopically from Groups 1 and 5, and target organs (liver andkidney) from Groups 2-4, by a veterinary pathologist certified by theAmerican College of Veterinary Pathologists.

Control vehicle included Peg-400 (Sigma P-3265), Tween® 80 (SigmaP-8074) and PVP-K30 (polyvinyl pyrrolidone or povidone K-30, Spectrum PNP1454) at a ratio of 90:5:5, w/w/w. Appropriate amounts of Compound 102were added to the control vehicle to produce homogeneous dosingsolutions/suspensions.

Results

Once daily, oral gavage administration of Compound 102 for 14 days toSprague-Dawley rats at 10, 100, 300 or 700 mg/kg/day was not associatedwith test article-related morbidity or early death. There were only two700 mg/kg/day female animals that died early on Days 3 and 9, but grossand microscopic findings indicate that each death was the result ofesophageal perforation during dosing.

Clinical observation data indicated an increased incidence and frequencyin rough hair coat and nasal discharge for animals dosed with Compound102 (particularly for the female animals at ≧100 mg/kg/day). Male andfemale animals dosed with Compound 102 exhibited an increased weightgain compared to control animals, which was an anticipatedpharmacological effect of this class of compounds.

Clinical pathology alterations attributed to administration of Compound102 included minimal changes in alkaline phosphatase (ALP), phosphorus,triglycerides, potassium, reticulocyte counts, red cell distributionwidth, platelet counts and mean platelet volume at doses ≧10 mg/kg/day.A minimal prolongation in prothrombin time was also noted for male andfemale animals dosed with Compound 102 at 300 and 700 mg/kg/day. Becauseof the small magnitude of change, none of these alterations wereconsidered to be adverse. There were no test article-related findings inthe urinalysis data. There were no macroscopic or microscopic changesthat were considered to be related to Compound 102. Dose-dependent liverweight increases were not correlated with appreciable increases inhepatocellular size or liver pathology.

On Day 1, systemic exposure of Compound 102 increased with increasingdose up to 300 mg/kg/day in both male and female rats. At 700 mg/kg/day,Compound 102 systemic exposure was not increased, indicating thatabsorption was saturated. There was a noticeable sex difference in Day 1toxicokinetic samples, with higher systemic exposure in females acrossall dose levels. Repeat administration of Compound 102 decreasedsystemic exposure, and the decrease in systemic exposure was morepronounced in the high dose groups.

Serum Chemistry

In female rats dosed at ≧10 mg/kg/day alkaline phosphatase values andphosphorus concentrations were minimally higher. These changes were notaccompanied by increases in bilirubin or GGT. Due to the alterationsnoted in phosphorus and ALP, it appears that Compound 102 may have aneffect on bone; however, no histological changes were noted. Thus, whilethese changes are attributed to administration of Compound 102, they arenot considered to be adverse, and may reflect the anabolic nature ofCompound 102.

Triglyceride concentrations were minimally lower in male rats dosed at≧100 mg/kg/day and in female rats dosed at ≧300 mg/kg/day (changes werestatistically significant). Potassium levels were minimally higher inmale rats dosed at ≧300 mg/kg/day and in female rats dosed at ≧100mg/kg/day. These minimal changes in potassium and triglycerides were notconsidered to be adverse. Intergroup differences in other serumchemistry parameters were sporadic or of a magnitude of change commonlyobserved in laboratory rats undergoing similar study procedures and werenot considered to the test article-related.

Hematology

Female rats dosed with ≧10 mg/kg/day Compound 102 had higherreticulocyte counts and red cell distribution width. These changes werenot accompanied by decreased indicators of circulating erythrocyte mass(i.e., red blood cells, hemoglobin or hematocrit) nor were the changesnoted in male animals. Platelet counts also were increased withstatistical significance for female animals dosed at ≧100 mg/kg/day andfor males dosed at 10 mg/kg/day and 700 mg/kg/day. Increases in meanplatelet count also were noted for the female animals (statisticallysignificant for Groups 3 and 5). The test article increases inreticulocyte counts, RDW, platelets and MPV were minimal and notconsidered to be adverse. Intergroup differences in the hematologychemistry parameters were sporadic or of a magnitude of change commonlyobserved in laboratory rats undergoing similar study procedures and werenot considered to the test article-related.

Coagulation

A minimal prolongation in prothrombin time was noted for male and femaleanimals dosed with Compound 102 at 300 and 700 mg/kg/day. The magnitudeof this change was not considered to be adverse.

Urinalysis

There were no alterations in the urinalysis parameters that wereattributed to administration of Compound 102.

Postmortem Observations

There were no macroscopic or microscopic changes that were consideredrelated to Compound 102. Dose-dependent liver weight increases were notcorrelated with appreciable increases in hepatocellular size and theliver pathology was negligible. There were, compared to concurrentcontrols, several statistically significant increases in group meanabsolute organ weights. Those absolute organ weights with dose-dependentincreases were limited to the liver of both sexes. The group mean bodyweights were increased compared to the controls at every dose level.Despite the increased body weights, liver to body weight ratios wereincreased in a doe-dependent manner for males and females at 100mg/kg/day and above (statistically significant at 300 and 700mg/kg/day). Liver to brain weight ratios also were increased in a dosedependent manner.

Conclusions

Once daily oral gavage administration of Compound 102 for 14 days toSprague-Dawley rats at 10, 100, 200 or 700 mg/kg/day was not associatedwith test article-related moribundity or early death. A statisticallysignificant increase in food consumption was noted for female animalsdosed with Compound 102 (all dose levels), which correlates with anincreased weight gain in these animals. No such change was noted formale animals. A dose independent test article-related increase in bodyweight occurred for male and female animals (of statistical significancebetween Days 8 and 14). The mean body weight gain in the high dose groupappeared less than that noted in the lower dose groups for male andfemale animals. There were no macroscopic or microscopic changes thatwere considered to be related to Compound 102. Dose-dependent liverweight increases were not correlated with appreciable increases inhepatocellular size or liver pathology.

Test article-related clinical signs included increased observations ofrough hair coat and nasal discharge for animals dosed at ≧100 mg/kg/day.Increases in body weight also were observed for male and female animalsdosed with Compound 102. Non-adverse clinical pathology alterationsrelated to administration of ≧10 mg/kg/day Compound 102 were noted foralkaline phosphatase, phosphorus, triglyceride, potassium, reticulocytecounts, red cell distribution width, platelet counts, mean plateletvolume and prothrombin time.

On Day 1, systemic exposure of Compound 102 increased with increasingdose up to 300 mg/kg/day in both male and female rats. At 700 mg/kg/day,Compound 102 showed a saturated absorption without further increase ofsystemic exposure. There was a noticeable sex difference in Day 1toxicokinetic sample, with higher systemic exposure in females acrossall dose levels. Repeat administration of Compound 102 decreasedsystemic exposure, and the decrease in systemic exposure was morepronounced in the high dose groups.

Based on the overall findings, and as the clinical observations of roughhair coat and nasal discharge (indications of stress) noted for animalsdosed at ≧100 mg/kg/day were not associated with additional clinicalfindings, the No-observed-adverse-effect level (NOAEL) for this studywas considered to be 700 mg/kg/day.

Example 19 14-Day Oral Toxicity and Toxicokinetic Study in CynomolgusMonkeys

A study was performed to determine any potential toxicity and thetoxicokinetic profile of Compound 102 when administered orally (vianasal gavage) to cynomolgus monkeys for at least 14 days. The cynomolgusmonkey was chosen for this study as it is a non-rodent species that iscommonly used for non-clinical toxicity evaluations. Use of the monkeymodel maximized the likelihood of identifying toxicological responsesthat may occur in humans. Ten experimentally naïve cynomolgus monkeys (5male and 5 female) from about 2.5 to 3 years of age and weighing betweenabout 2.3 to about 2.8 kg at Day −1 of the study were assigned to dosegroups as shown in Table 6 below.

TABLE 6 Dose Groups. No. of Dose No. Group Males/ Dose Level Volume DoseSolution Necropsied No. Females (mg/kg) (mL/kg) Conc. (mg/ml) on Day 151 1/1  0 (control) 2 0 1/1 2 1/1  5 2 2.5 1/1 3 1/1  50 2 25 1/1 4 1/1150 2 75 1/1 5 1/1 450 2 225 1/1All animals were dosed via nasal gavage once daily for 14 consecutivedays. The first day of dosing was designated Day 1 for all animals. Theanimals were evaluated for clinical signs (evaluation of mortality andmorbidity, twice daily), cage side observations and food consumption(once daily), body weight (Days 1, 8 and 14), physical examination (Day−5 and post dosing following the 13/24 hour toxicokinetic collection onDay 14), ophthalmic examination (prestudy and owing the Day 13/24 hourtoxicokinetic collection on Day 14), and clinical pathology parameters(serum chemistry, hematology and coagulation on Days −3 and 14), urinesamples were collected during necropsy (Day 15) for urinalysis. Bloodsamples were collected for toxicokinetic analysis at 1, 2, 4, 8, 12 and24 hours post dose on Days 1 and 13.

All ten animals were euthanized one day after the last dose. Attermination, a full necropsy was conducted on all animals, and tissueswere collected, preserved, processed and examined microscopically by anAmerican College of Veterinary Pathologists (ACVP) certifiedpathologist.

Vehicle was composed of PEG 400/Tween® 80/PVP-K30 (90/5/5, w/w/w).Appropriate amounts of Compound 102 were added to vehicle to prepare thesuspension of test compound administered to the animals.

Blood samples for evaluation of serum chemistry, hematology andcoagulation parameters were collected from all animals on Days −3 and14. Urine samples were obtained by bladder puncture during necropsy. Theanimals were fasted for at east 8 hours prior to blood collections forserum chemistry. Serum chemistry parameters included alanineaminotransferase (ALT), total protein, aspartate aminotransferase (AST),alkaline phosphatase (ALP), gamma-glutamyltransferase (GGT), albumin,globulin, albumin/globulin ratio, lactate dehydrogenase (LD), glucose,total bilirubin, cholesterol, urea nitrogen (BUN), creatinine,triglycerides, sodium, calcium, potassium, phosphorus, chloride andcarbon dioxide. Hematology parameters included red blood cell count,hemoglobin concentration, hematocrit, mean corpuscular volume (MCV),mean corpuscular hemoglobin concentration (MCHC), mean corpuscularhemoglobin (MCH), red cell distribution width, reticulocyte count,platelet count, mean platelet volume (MPV) and white blood cell count.Coagulation parameters included prothrombin time (PT) and activatedpartial thromboplastin time. Urinalysis parameters includedcolor/character, specific gravity, pH, protein, glucose, ketones,bilirubin, occult blood and microscopics.

Results

All animals survived until scheduled necropsy. There were no Compound102-related clinical signs or effects on food consumption. Watery feceswas observed frequently for all study animals beginning on Day 1. Thewatery feces were attributed to the PEG400 component of the vehicle,which is known to produce gastrointestinal disturbances, includingdiarrhea, following ingestion.

There was a dose-independent increase in body weight in all of theCompound 102-dosed animals relative to controls. The magnitude of theincreases to individual animal body weight during the study wasconsidered notable and consistent with the expected pharmacologicalactivity of the class of compound (i.e., SARMs are known to beandrogenic and increase bone and muscle mass; see Shalender et al.,Nature Clinical Practice Endocrinology & Metabolism 2: 146-159 (2006)).There were no Compound 102-related findings identified during ophthalmicexaminations.

Serum Chemistry

Serum cholesterol concentration was decreased on Day 14 in all animalsreceiving Compound 102. The reductions in cholesterol did not appear tobe adverse. Ater 14 days of dosing, the female animal dosed with 450mg/kg had elevated serum alanine aminotransferase (ALT) and aspartateaminotransferase (AST) activity, without increased lactate dehydrogenase(LD). This animal also had barely noticeable minimal single cellhepatocellular necrosis in the liver. Taken together, these changesaffecting a high-dose animal suggested a possible relationship with thetest article. Other changes in serum chemistry parameters includingsmall increases in gamma-glutamyltransferase on Day 14 in severalanimals in dosed groups were attributed to biological variabilitycommonly observed in laboratory-housed cynomolgus monkeys undergoingsimilar study procedures.

Hematology

There was a dose-independent decrease in platelet count at all Compound102 dose levels. On Day 14, platelet counts were lower (approximately14% to 38%) relative to Day −3 in both male and female animals in the 5,50 and 150 mg/kg-dosed groups and the female high-dose animal (450mg/kg). Platelet count on Day 14 was marginally increased for thehigh-dose and control males and reduced approximately 5% for the controlfemale relative to Day −3. Although the platelet counts were lower for 7of 8 Compound 102-dosed animals on Day 14 (relative to Day −3) and tothe control animals, the clinical significance of this finding was notconsidered adverse.

The indicators of circulating erythrocyte mass were mildly reduced onDay 14 (relative to Day −3) in all 5 groups, due to toxicokinetic bloodsample collection on Day 13. Red cell distribution width andreticulocyte counts were mildly increased on Day 14 (relative to Day −3)in all 5 groups; these changes reflect regenerative erythropoiesisresulting from blood collections on Day −3 (for clinical pathology) andDay 1 (for toxicokinetics). Fluctuations in total leukocyte (WBC),neutrophil, lymphocyte and monocyte counts which were most consistent inboth Group 4 animals were dose-independent and likely incidental.

Coagulation

On Day 14, all animals in Compound 102-dosed groups, except for the 5mg/kg-dosed female, had minimally prolonged (1.8 to 3.5 seconds longerthan Day −3) prothrombin time (PT); PT in the control animals wassimilar to Day −3. The changes did not exhibit a dose response and werenot considered adverse.

Urinalysis

There were no Compound 102-related effects on urinalysis parameters.

Macroscopic Observations

Gross necropsy findings suggestive of a relationship to Compound 102administration were limited to decreased thymus size and browndiscoloration of the adrenals in both sexes at 150 and 450 mg/kg.Decreased thymus size correlated microscopically with minimal to mildlymphoid depletion on the thymic cortex. There was no microscopiccorrelate to the brown adrenal discoloration.

Histopathology

There were no direct Compound 102-related microscopic findings. Minimalto mild lymphoid cortical depletion of the thymus was present in malesat 150 mg/kg and above and at 50 mg/kg and above in females. Thisobservation, however, was considered secondary to stress and not adirect toxicologic effect. Most microscopic findings were randomlydistributed in control and dosed animals or were considered commonincidental findings in cynomolgus monkeys and not believed to be relatedto Compound 102 dosing.

Conclusions

Daily administration of 5, 50, 150 and 50 mg/kg Compound 102 for 14 daysdid not result in mortality, adverse clinical signs or changes in foodconsumption or histologic effects. There were no abnormal findingsidentified during physical or ophthalmic examinations, or fromurinalysis.

Compound 102-related effects were identified at the 5, 50, 150 and 450m/kg dose levels in both sexes, including increased body weight(consistent with the expected pharmacological activity of this class ofcompounds), decreased serum cholesterol concentration and plateletcount, and prolonged prothrombin time. These changes were generallydose-independent and considered non-adverse.

Macroscopic (gross necropsy) observations possibly related to Compound102 were limited to brown discoloration of the adrenals and decreasedthymus size at 150 and 450 mg/kg in both sexes. There was no apparentmicroscopic correlate to the brown adrenals. Minimal to mild lymphoidcortical depletion of the thymus in males at 150 mg/kg and above and infemales at 50 mg/kg and above and decreased thymus size were consideredsecondary to stress and not a direct toxicologic effect.

The systemic exposure of Compound 102 increased with increasing dose upto 450 mg/kg in both sexes. Following repeat administration of Compound102 the systemic exposure was decreased on Day 13 (relative to Day 1exposure), and the decrease in systemic exposure was more pronounced inthe high-dose groups. There was no noticeable sex difference in the Day1 and Day 13 toxicokinetic parameters.

Under the conditions of this study, the NOAEL was 450 mg/kg for themales and 150 mg/kg for the females (based on findings that in the 450mg/kg-dosed female ALT and AST were elevated and there were histologicchanges to the liver). A NOEL was not achieved as a result of increasedbody weight, decreased serum cholesterol concentration and plateletcount, and prolonged prothrombin time present in all Compound 102-dosedgroups.

Since modifications will be apparent to those of skill in this art, itis intended that this invention be limited only by the scope of theappended claims.

The invention claimed is:
 1. A method for modulating an androgenreceptor, comprising: contacting an androgen receptor with a compound offormula I:

wherein: R¹ is CF₃, F or Cl; R² is H or methyl; and R³ is H or methyl,or a pharmaceutically acceptable salt or ester thereof.
 2. The method ofclaim 1, wherein the androgen receptor is activated by the compound. 3.The method of claim 1, wherein the androgen receptor is deactivated bythe compound.
 4. The method of claim 1, wherein the compound of formulaI is a selective androgen receptor agonist.
 5. The method of claim 1,wherein the compound of formula I is a selective androgen receptorpartial agonist.
 6. The method of claim 1, wherein the compound offormula I is a tissue-specific androgen receptor agonist.
 7. The methodof claim 1, wherein the androgen receptor is in a cell.
 8. The method ofclaim 1, wherein R¹ is CF₃.
 9. The method of claim 8, wherein R² is H.10. The method of claim 9, wherein R³ is H.
 11. The method of claim 1,wherein R² is H.
 12. The method of claim 11, wherein R³ is H.
 13. Themethod of claim 1, wherein R³ is H.
 14. The method of claim 1, whereinthe compound of formula I is selected from among:R,R-4-(2-(1-Hydroxyl-2,2,2-trifluoroethyl)pyrrolidinyl)-2-trifluoromethyl-benzonitrile;4-(2(R)-(1(S)-hydroxyl-2,2,2-trifluoroethyl)pyrrolidinyl)-2-trifluoromethyl-benzonitrile;R,R,R-4-(2-(1-Hydroxyl-2,2,2-trifluoroethyl)-5-methylpyrrolidinyl)-2-trifluoromethyl-benzonitrile;4-(2(R)-(1(S)-hydroxyl-2,2,2-trifluoroethyl)-5(R)-methylpyrrolidinyl)-2-trifluoromethyl-benzonitrile;R,R-4-(2-(1-Hydroxyl-2,2,2-trifluoroethyl)pyrrolidinyl)-2-chlorobenzonitrile;4-(2(R)-(1(S)-hydroxyl-2,2,2-trifluoroethyl)pyrrolidinyl)-2-chlorobenzonitrile;R,R,R-4-(2-(1-hydroxyl-2,2,2-trifluoroethyl)-5-methylpyrrolidinyl)-2-chlorobenzonitrile;4-(2(R)-(1(S)-hydroxyl-2,2,2-trifluoroethyl)-5(R)-methylpyrrolidinyl)-2-chlorobenzonitrile;R,R-4-(2-(1-hydroxyl-2,2,2-trifluoroethyl)pyrrolidinyl)-2-chloro-3-methylbenzonitrile;4-(2(R)-(1(S)-hydroxyl-2,2,2-trifluoroethyl)-pyrrolidinyl)-2-chloro-3-methylbenzonitrile;3-methyl-4-((R)-2-((R)-2,2,2-trifluoro-1-hydroxyethyl)pyrrolidin-1-yl)-2-(trifluoromethyl)-benzonitrile;3-methyl-4-((R)-2-((S)-2,2,2-trifluoro-1-hydroxyethyl)pyrrolidin-1-yl)-2-(trifluoromethyl)-benzonitrile;3-methyl-4-((2R,5R)-2-methyl-5-((S)-2,2,2-trifluoro-1-hydroxyethyl)-pyrrolidin-1-yl)-2-(trifluoromethyl)benzonitrile;2-fluoro-4-((2R,5R)-2-methyl-5-((S)-2,2,2-trifluoro-1-hydroxyethyl)-pyrrolidin-1-yl)-benzonitrile;2-fluoro-3-methyl-4-((2R,5R)-2-methyl-5-((S)-2,2,2-trifluoro-1-hydroxyethyl)-pyrrolidin-1-yl)benzonitrile;2-fluoro-3-methyl-4-((R)-2-((S)-2,2,2-trifluoro-1-hydroxyethyl)pyrrolidin-1-yl)-benzonitrile;2-fluoro-4-((R)-2-((S)-2,2,2-trifluoro-1-hydroxyethyl)pyrrolidin-1-yl)benzonitrile;2-chloro-4-((2R,5R)-2-methyl-5-((S)-2,2,2-trifluoro-1-hydroxyethyl)-pyrrolidin-1-yl)-benzonitrile;2-chloro-3-methyl-4-((2R,5R)-2-methyl-5-((S)-2,2,2-trifluoro-1-hydroxyethyl)-pyrrolidin-1-yl)benzonitrile;2-chloro-3-methyl-4-((R)-2-((S)-2,2,2-trifluoro-1-hydroxyethyl)pyrrolidin-1-yl)-benzonitrile;and2-chloro-4-((R)-2-((S)-2,2,2-trifluoro-1-hydroxyethyl)pyrrolidin-1-yl)-benzonitrile;or a pharmaceutically acceptable salt thereof.
 15. The method of claim1, wherein the compound of formula I is4-(2(R)-(1(S)-hydroxyl-2,2,2-trifluoroethyl)pyrrolidinyl)-2-trifluoromethyl-benzonitrile,or a pharmaceutically acceptable salt thereof.